IMMUNOSERO POST TEST (03/15/2025)

1. Which of the ff is considered as a primary antigen recognition protein of the innate immune
system
A.​ Complement
B.​ Antibodies
C.​ МНС
D.​ TLRs
RATIONALE:
A.​ It does not recognize antigens in the same way as receptors like TLRs.
B.​ Antibodies (Immunoglobulins) are part of the adaptive immune system, not
innate immunity.
C.​ They do not function as primary antigen recognition proteins in the innate
immune system.
D.​ TLRs are pattern recognition receptors (PRRs) of the innate immune
system. They detect pathogen-associated molecular patterns (PAMPs) found
on microbes (e.g., bacterial lipopolysaccharides, viral RNA).
Reference: Akira, S., Uematsu, S., & Takeuchi, O. (2006). "Pathogen recognition and
innate immunity." Cell, 124(4), 783-801.

2. What type of immunity develops during convalescence from an infection?

A.​ natural active immunity
B.​ natural passive immunity
C.​ artificial active immunity
D.​ artificial passive immunity
RATIONALE:
A.​ Develops after an individual recovers (convalesces) from an infection because
their immune system has encountered the pathogen, mounted a response, and
formed immunological memory (B and T cell activation).
B.​ Occurs when antibodies are transferred naturally from one individual to
another without immune system activation
C.​ Involves vaccination, where an individual is exposed to a weakened, killed, or
component of a pathogen, triggering an immune response.
D.​ Involves the direct administration of pre-formed antibodies (e.g., monoclonal
antibodies, immune globulin therapy).
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

3. Major humoral component of innate immunity

A.​ Complement
B.​ Antibodies
C.​ Interferons
D.​ Interleukins
RATIONALE:
A.​ The complement system is the major humoral (soluble) component of the innate
immune system.
B.​ Antibodies (immunoglobulins) are humoral components of adaptive
immunity, not innate immunity.
C.​ While they are part of the innate immune response, they function
intracellularly rather than as a humoral (circulating) component like
complement.
D.​ They regulate inflammation but are not major humoral components like
complement.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

4. Which of the ff components of the immune system may be increased within 72 hrs after an
initial infection
A.​ Neutrophils, antibody, complement, acute-phase reactants
B.​ Macrophages, T cells, antibody, haptoglobin, fibrinogen
C.​ Neutrophils, macrophages, complement, fibrinogen, C-reactive protein
D.​ Macrophages, T cells, B cells, ceruloplasmin, complement
RATIONALE:
A.​ Antibodies, however, take longer to develop (typically 5–7 days for IgM, and
weeks for IgG), so this answer is incorrect.
B.​ However, T cells and antibodies are part of the adaptive immune response,
which typically takes several days to weeksto activate, making this answer
incorrect.
C.​ Fibrinogen & C-reactive protein (CRP) – Acute-phase proteins produced by
the liver in response to inflammation, peaking within 24–72 hours.
D.​ T cells and B cells belong to the adaptive immune response, which takes
several days to weeks to activate, making this option incorrect.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

5. Process by which phagocytic cells are attracted toward an area where they detect a
disturbance in the normal functions of
A.​ Diapedesis
B.​ Opsonization
C.​ Sensitization
D.​ Chemotaxis
RATIONALE:
A.​ Diapedesis (extravasation or transmigration) is the process by which immune
cells (e.g., neutrophils, monocytes) move through the endothelial wall of blood
vessels to enter tissues.
 B.​ Opsonization is the process by which pathogens are coated with opsonins
(e.g., IgG, C3b) to enhance phagocytosis.
C.​ Sensitization refers to the priming of immune cells for a stronger response
upon subsequent exposure.
D.​ Chemotaxis is the process by which immune cells (e.g., neutrophils,
macrophages) are attracted to the site of infection or tissue damage in
response to chemical signals.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

6. All of the following serum proteins are elevated in acute-phase reactions or inflammation,
except:
A.​ Ceruloplasmin
B.​ Transferrin
C.​ Haptoglobin
D.​ Alpha-1 antitrypsin
RATIONALE:
A.​ Ceruloplasmin is an acute-phase reactant that increases during inflammation.
B.​ Transferrin is not an acute-phase reactant; instead, its levels decrease during
inflammation (negative acute-phase protein).
C.​ Haptoglobin is a positive acute-phase reactant that binds free hemoglobin to
prevent oxidative damage.
D.​ AAT is a major acute-phase protein that inhibits proteases, particularly
neutrophil elastase, to prevent tissue damage.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

7. Select the correct order of maturation sequence for T-cells:

A.​ BM → Thymic medulla → Thymic cortex → peripheral lymphoid organs
B.​ BM → Thymic cortex → Thymic medulla → peripheral lymphoid organs
C.​ BM → peripheral lymphoid organs → Thymic cortex → Thymic medulla
D.​ Peripheral lymphoid organs → BM → Thymic medulla → Thymic cortex Cortex; primary
follicles
RATIONALE:
A.​ T-cell precursors originate in the bone marrow (BM) but do not mature there.
B.​ T-cell precursors originate in the BM and travel to the thymus for maturation.
C.​ T-cell precursors do not mature in peripheral lymphoid organs before thymic
development.
D.​ T-cell precursors do not originate in peripheral lymphoid organs.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.
8. B lymphocytes account for about 10% to 20% of all lymphocytes in the blood; they are also
found in_of lymph nodes and within_ in the spleen.
A.​ Cortex; primary follicles
B.​ Cortex; periarteriolar lymphoid sheath
C.​ Paracortex; primary follicles
D.​ Paracortex; periarteriolar lymphoid sheath
RATIONALE:
A.​ B lymphocytes (B cells) are primarily found in the cortex of lymph nodes. B cells
also localize within primary follicles in the white pulp.
B.​ The cortex of lymph nodes contains B cells, but PALS is found in the
spleen, not lymph nodes.
C.​ The paracortex of lymph nodes is T-cell rich, while B cells are mainly in the
cortex and follicles.
D.​ The paracortex of lymph nodes is where T cells reside, not B cells.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

9. Which surface markers are found on mature, peripheral helper T cells?

A.​ CD1, CD2, CD4
B.​ CD2, CD3, CD8
C.​ CD1, CD3, CD4
D.​ CD2, CD3, CD4
RATIONALE:
A.​ CD1 is involved in lipid antigen presentation and is mainly expressed by
antigen-presenting cells (APCs), not mature helper T cells.
B.​ CD8 is a marker of cytotoxic T cells (CTLs), not helper T cells.
C.​ CD1 is not a marker of helper T cells; it is found on APCs and plays a role in
lipid antigen presentation.
D.​ All are found on T cells such as CD4 on helper T cells (Th cells), which assist in
activating B cells, macrophages, and cytotoxic T cells.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

10. Which surface markers are found on plasma cells?

A.​ CD38 and CD138
B.​ CD16 and CD56
C.​ CD2 and CD3
D.​ CD33 and CD34
RATIONALE:
A.​ A marker of plasma cell differentiation and antibody-secreting activity and
hallmark marker of plasma cells, involved in cell adhesion and extracellular
matrix interactions.
 B.​ Found on natural killer (NK) cells, monocytes, and neutrophils, associated
with antibody-dependent cellular cytotoxicity (ADCC).
C.​ An adhesion molecule found on T cells and NK cells, aiding in T-cell
activation.
D.​ A marker of myeloid lineage cells, commonly expressed on monocytes,
macrophages, and myeloid progenitors.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

11. Frequently used for immunologic identification of B cells:

A.​ CD16
B.​ CD19
C.​ CD21
D.​ CD25
RATIONALE:
A.​ CD16 (FcγRIII) is a receptor for the Fc portion of IgG.
B.​ CD19 is a pan-B cell marker, expressed from early B-cell development until
plasma cell differentiation.
C.​ CD21 (Complement Receptor 2, CR2) is expressed on mature B cells and
follicular dendritic cells.
D.​ CD25 (IL-2 receptor alpha chain) is a marker of regulatory T cells (Tregs) and
activated T cells, not B cells.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

12. All are correct about cytotoxic T cells and NK cells, except:
A.​ release perforins and granzymes
B.​ effective against virally infected cells
C.​ cellular components of the immune system
D.​ do not exhibit MHC class restriction
RATIONALE:
A.​ Both cytotoxic T cells (CD8⁺ T cells) and natural killer (NK) cells eliminate infected
or malignant cells via perforin and granzyme-mediated apoptosis.
B.​ Cytotoxic T cells (CTLs) recognize viral peptides presented on MHC class I
molecules and induce apoptosis in infected cells.
C.​ Both CD8⁺ cytotoxic T cells and NK cells are cellular components of the
immune system, involved in cell-mediated immunity.
D.​ Cytotoxic T cells (CD8⁺ T cells) are MHC class I-restricted, meaning they
require antigen presentation by MHC class I molecules on infected or malignant
cells.
Reference:Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.
13. Which of the following found on the surface of most T cells recognizes antigen?
A.​ CD2
B.​ CD3
C.​ MHC proteins
D.​ TCR
RATIONALE:
A.​ CD2 is an adhesion molecule found on T cells and natural killer (NK) cells.
B.​ CD3 is a signaling molecule complex associated with the T-cell receptor
(TCR).
C.​ Major Histocompatibility Complex (MHC) proteins are found on
antigen-presenting cells (APCs) and other nucleated cells.
D.​ TCR is the antigen-specific receptor found on the surface of T cells.
Reference: Murphy, K., Weaver, C., & Berg, L. J. (2022). Janeway’s immunobiology (10th
ed.). Garland Science.

14. The ______ T cell can help B lymphocytes differentiate to plasma cells, or it can help other
T lymphocytes differentiate to either cytotoxic cells or cells with suppressor function.​
a. CD2+
b. CD3+
c. CD4+
d. CD8+

RATIONALE:
a.​ CD2+ – CD2 is a surface marker involved in T-cell adhesion and signaling but not
specifically responsible for helper functions.
b.​ CD3+ – CD3 is part of the T-cell receptor complex found on all T cells; it doesn't indicate
helper or cytotoxic function.
c.​ CD4+ – CD4+ T cells are helper T cells that assist B cells in producing antibodies and
help activate other immune cells, including CD8+ cytotoxic T cells.
d.​ CD8+ – CD8+ T cells are primarily cytotoxic and do not help in B cell differentiation.

15. Function of memory cells:

a.​ for the manufacture of specific antibodies

b.​ for antigenic stimulation
c.​ for immune tolerance
d.​ for anamnestic response

RATIONALE:

a.​ for the manufacture of specific antibodies – This is the role of plasma cells, not memory
cells.
b.​ for antigenic stimulation – Antigenic stimulation activates naive cells but is not a function
of memory cells.
 c.​ for immune tolerance – Immune tolerance is associated with regulatory cells, not
memory cells.
d.​ for anamnestic response – Memory cells respond rapidly and strongly upon re-exposure
to the same antigen, resulting in a quicker immune response.

16. Which MHC class of molecule is necessary for antigen recognition by Th cells?

a.​ Class I
b.​ Class II
c.​ Class III
d.​ No MHC molecule is necessary for antigen recognition

RATIONALE:

a.​ Class I – Class I MHC presents antigen to CD8+ cytotoxic T cells, not helper T cells.
b.​ Class II – Class II MHC presents antigen to CD4+ helper T cells, allowing their
activation.
c.​ Class III – Class III MHC encodes proteins like complement components, not
antigen-presenting molecules.
d.​ No MHC molecule is necessary for antigen recognition – MHC is essential for antigen
recognition by T cells.

17. Presentation of antigen by class I molecules allows T cells to detect and mount a
_________ response to a foreign material

a.​ Phagocytic
b.​ Cytotoxic
c.​ Complement-mediated
d.​ Humoral-mediated

RATIONALE:

a.​ Phagocytic – Phagocytosis is performed by macrophages and neutrophils, not T cells.

b.​ Cytotoxic – Class I MHC presents antigens to CD8+ cytotoxic T cells, which then kill
infected or abnormal cells.
c.​ Complement-mediated – Complement is part of innate immunity, not directly linked with
MHC class I presentation.
d.​ Humoral-mediated – Humoral responses involve B cells and antibodies, not CD8+ T
cells.

18. What type of cells are usually used in clinical HLA Class II phenotyping?

a.​ erythrocytes
b.​ stem cells
 c.​ purified T cells
d.​ purified B cells

RATIONALE:

a.​ erythrocytes – Red blood cells do not express HLA class II molecules.
b.​ stem cells – Stem cells are not typically used in routine HLA phenotyping.
c.​ purified T cells – T cells express class I MHC but not class II.
d.​ purified B cells – B cells express class II MHC molecules and are ideal for phenotyping.

19. Which MHC class has chains structure of a - chain and beta 2 - microglobulin?

a.​ I
b.​ II
c.​ III
d.​ none of the above

RATIONALE:

a.​ I – MHC class I molecules consist of a heavy alpha chain and a β2-microglobulin light
chain.
b.​ II – MHC class II is made of two chains: alpha and beta, both membrane-anchored.
c.​ III – MHC class III does not refer to a structured complex but includes genes for
complement and cytokines.
d.​ none of the above – MHC class I is the correct match for this structural description.

20. Which disease is associated with inheritance of HLA-DQ2, DQ8?

a.​ Ankylosing spondylitis

b.​ Gluten-sensitive enteropathy
c.​ Rheumatoid arthritis
d.​ Myasthenia gravis

RATIONALE:

a. Ankylosing spondylitis – Strongly associated with HLA-B27, not DQ2/DQ8.

b. Gluten-sensitive enteropathy – Also known as celiac disease, it has a strong genetic link with
HLA-DQ2 and DQ8.

c. Rheumatoid arthritis – Typically associated with HLA-DR4, not DQ2/DQ8.

d. Myasthenia gravis – More often associated with HLA-DR3, not DQ2/DQ8.

21. Immunoglobulin variation which refers to antigenic individuality specific to each Ig molecule:

a.​ Isotype
b.​ Idiotype
c.​ Allotype
d.​ A and C

RATIONALE:

a. Isotype – Incorrect. Isotype refers to the class and subclass variations of immunoglobulins
(e.g., IgG, IgA), which are constant across all individuals of a species.

b. Idiotype – Correct. Idiotype refers to the unique set of antigenic determinants located in the
variable region of the antibody that gives it specificity. These determinants are unique to each
antibody clone and define its antigen-binding site.

c. Allotype – Incorrect. Allotype refers to genetic variations within constant regions of

immunoglobulins that vary between individuals of the same species.

d. A and C – Incorrect. Although isotype and allotype are valid forms of immunoglobulin
variation, neither refers to the unique antigen specificity found in idiotypes.

22. IgM molecule is considered _____ and its J-chain is an acidic glycoprotein with a high
content of _____ residues

a.​ Bivalent; proline

b.​ Tetravalent; proline
c.​ Pentavalent; cysteine
d.​ Decavalent; cysteine

RATIONALE:

a. Bivalent; proline – Incorrect. IgM is not bivalent; that refers to IgG and IgE, which have two
antigen-binding sites.

b. Tetravalent; proline – Incorrect. Tetravalency isn't a feature of IgM; IgA dimers may have four
binding sites.

c. Pentavalent; cysteine – Incorrect. While IgM has five monomeric units, it has 10 binding sites,
making it decavalent.

d. Decavalent; cysteine – Correct. IgM has five monomers joined by a J-chain, making it
decavalent. The J-chain contains multiple cysteine residues that facilitate disulfide bonding,
ensuring stability and polymerization.
23. Antibodies which are principally bound to cells: 1. IgG 2. IgD 3. IgE 4. IgA​

a.​ 1 and 2
b.​ 1 and 3
c.​ 2 and 3
d.​ 3 only

RATIONALE:

a. 1 and 2 – Incorrect. IgG is mainly found in circulation and tissues, not principally bound to
cells.

b. 1 and 3 – Incorrect. While IgE binds to mast cells and basophils, IgG is not cell-bound by
default.

c. 2 and 3 – Correct. IgD is primarily found on the surface of immature B cells, while IgE binds
tightly to receptors on mast cells and basophils.

d. 3 only – Incorrect. IgE is cell-bound, but this choice ignores IgD, which also exhibits
cell-binding behavior on B cells.

24. Antibodies which can fix properdin complement pathway: 1. IgG 2. IgM 3. IgA 4. IgE

a.​ 1 and 2
b.​ 1 and 3
c.​ 2 and 3
d.​ 3 only

RATIONALE:

a. 1 and 2 – Incorrect. IgG and IgM activate the classical complement pathway, not the
properdin (alternative) pathway.

b. 1 and 3 – Incorrect. While IgA participates in the alternative pathway, IgG is only involved in
the classical pathway.

c. 2 and 3 – Incorrect. IgM does not activate the properdin pathway; it initiates the classical
pathway.

d. 3 only – Correct. Only IgA, particularly in its secretory form, can activate the alternative
(properdin) complement pathway. This activation does not require antibodies to bind to antigen
first and does not involve C1, unlike the classical pathway.

25. Which of the following antibody domains has a function of binding neutrophils?
 a.​ CH1
b.​ CH2
c.​ CH3
d.​ CH4

RATIONALE:

a. CH1 – Incorrect. The CH1 domain helps in stabilizing the variable region and contributes to
antigen binding but does not interact with immune effector cells.

b. CH2 – Incorrect. The CH2 domain is involved in complement binding, particularly in IgG and
IgM, but not directly in neutrophil interaction.

c. CH3 – Correct. The CH3 domain is part of the Fc region of IgG antibodies and is responsible
for binding to Fc gamma receptors (FcγRs) on neutrophils. This interaction enables immune
functions like phagocytosis and ADCC (antibody-dependent cellular cytotoxicity).

d. CH4 – Incorrect. CH4 is found only in IgM and IgE and does not play a role in neutrophil
binding.

26. Which antibody inhibits the adherence of microorganisms to the surface of mucosal
cells, preventing their entry into body tissues?​
a. IgA

b. IgM

c. IgD

d. IgE

RATIONALE:

a. IgA – Correct. Secretory IgA is the main antibody found in mucosal secretions (e.g., saliva,
tears, mucus) and acts as the first line of defense by blocking microbial adherence to epithelial
cells, thus preventing invasion.

b. IgM – Incorrect. IgM is primarily involved in early systemic immune responses and
complement activation, not mucosal protection.

c. IgD – Incorrect. IgD mainly functions as a receptor on naïve B cells and has limited direct
antimicrobial activity.

d. IgE – Incorrect. IgE is involved in allergic reactions and defense against parasites, not
mucosal barrier protection.
27. All of the following are functions of immunoglobulins except:​
a. Neutralizing toxic substances

b. Facilitating phagocytosis through opsonization

c. Interacting with Tc cells to lyse viruses

d. Combines with complement to destroy antigens

RATIONALE:

a. Neutralizing toxic substances – Correct function. Antibodies bind and neutralize toxins,
preventing them from affecting host cells.

b. Facilitating phagocytosis through opsonization – Correct function. Antibodies coat pathogens,

making them more recognizable to phagocytes for destruction.

c. Interacting with Te cells to lyse viruses – Incorrect function. Cytotoxic T cells (Te), not
antibodies, directly lyse virus-infected cells. Antibodies do not interact with T cells for this
purpose.

d. Combines with complement to destroy antigens – Correct function. Certain immunoglobulin

classes (like IgG and IgM) activate the complement system to lyse or opsonize pathogens.

28. What is measured in the CH50 assay?

a. RBC quantity needed to agglutinate 50% of antibody

b. Complement needed to lyse 50% of RBCs

c. Complement needed to lyse 50% of antibedv-sensitized RBCs

d. Antibody and complement needed to sensitize 50% of RBCs

Answer:

A. RBC quantity is not the focus of the CH50 assay.

B. The CH50 assay specifically measures complement activity, not just the complement needed
to lyse RBCs without antibodies.
C. correct because it describes the key principle of the CH50 assay: it measures the
complement components required to lyse 50% of RBCs that have been sensitized with antibody.

D. This option mixes up the concept, as the CH50 assay specifically measures the complement
activity in the context of antibody-sensitized RBCs, not antibody and complement needed to
sensitize RBCs.

29. Complement activity of Classical, Alternative, Lectin pathway in cases of C2 inherited

deficiency, respectively.

a. Low; Normal: Low

b. Low; Normal; Normal

c. Normal; Low; Normal

d. Normal; Low; Low

Answer:

A. correct because: Classical pathway: Low (C2 is required, and it’s deficient). Alternative
pathway: Normal (C2 is not involved in this pathway). Lectin pathway: Low (C2 is also required
for this pathway).

B. Low; Normal; Normal is incorrect because the lectin pathway should be low, not normal.

C. Normal; Low; Normal is incorrect because the classical pathway should be low, not normal.

D. Normal; Low; Low is incorrect because the classical pathway should be low, not normal, and
the alternative pathway should be normal, not low.

30. Inherited deficiencies of C1, C2 and C4 will result to low titer of CH50 and hm titer of AH50.

a. Normal; Low

b. Normal; High

c. Low; Normal

d. High; Normal
ANSWER:

A. Normal; Low: CH50 should be low, not normal, due to the classical pathway deficiency.

B. Normal; High: CH50 should be low, not normal, and AH50 wouldn't be high, just normal.

C. Low; Normal: Deficiencies in C1, C2, or C4 result in low CH50 since the classical pathway
can't function properly.

D. High; Normal: CH50 should be low, not high, due to the deficiency in classical pathway
components.

31. What type of disorders would show a decrease in C3, C4, and CH50?

a. Autoimmune disorders such as SLE and RA

b. Common variable immunodeficiency

c. Inflammatory process associated with tissue injury

d. Bacterial, viral, fungal, or parasitic infections

ANSWER:

A. Autoimmune disorders such as SLE and RA: While these disorders do cause decreased C3,
C4, and CH50 due to complement activation and consumption, the best answer in this case is c
(inflammatory processes), as it encompasses a broader range of conditions that also lead to
complement consumption.

B. Common variable immunodeficiency: This condition primarily affects B cells and antibody
production. It does not generally cause a decrease in C3, C4, or CH50, unless there is a
specific, separate complement deficiency.

C. Inflammatory process associated with tissue injury is correct because it leads to activation
and consumption of complement proteins (C3, C4) and decreases in CH50 as part of the
immune response.

D. Bacterial, viral, fungal, or parasitic infections: While infections can activate the complement
system, they do not typically cause significant depletion of C3, C4, or CH50. Infections might
cause a transient increase in complement activity but not usually a marked decrease.
32. Cell-associated complement regulatory protein which inhibits formation of Membrane attack
complex:

a. CD55 and CD59

b. CR1 and MCP

c. CD59 and HRF

d. CD55 and S protein

ANSWER:

A. CD55 and CD59: CD55 accelerates decay of convertases but does not directly inhibit MAC
formation.

B. CR1 and MCP: These regulate complement activation but do not prevent MAC formation.

C. CD59 and HRF because both proteins directly inhibit the formation of the Membrane Attack
Complex (MAC) by preventing C9 binding to the C5b-8 complex.

D. CD55 and S protein: S protein prevents C5b-7 from inserting into the membrane, but does
not directly block MAC formation.

33. Complement Fixation test can be use to:

a. detect complement component deficiencies

b. detect viral, fungal, and rickettsial antibodies

c. evaluate classical and alternative pathway

d. measure individual complement fragments

ANSWER:

A. detect complement component deficiencies: The Complement Fixation test is not designed to
detect deficiencies in individual complement components. Tests like CH50 or specific
complement component assays (e.g., C3, C4) are used to assess complement deficiencies.

B. detect viral, fungal, and rickettsial antibodies is correct because the CFT is used to detect
specific antibodies against these pathogens.
C. evaluate classical and alternative pathway: The Complement Fixation test does not directly
evaluate the classical or alternative pathways of complement. It is more focused on detecting
the presence of antibodies, not on pathway-specific activities.

D. measure individual complement fragments: The Complement Fixation test does not measure
individual complement fragments (like C3a, C5a, etc.). Other specific tests, such as ELISA or
Western blot, are used for measuring complement fragments.

34. Cytokine which has anti-inflammatory properties, depress cytokine-mediated lethality and
downregulate autoimmune disease:

a. IL-1

c. TNF-alpha

b. IL-6

[Link]-10.

ANSWER:

A. IL-1: IL-1 is a pro-inflammatory cytokine involved in promoting inflammation, fever, and

immune responses. It plays a role in several inflammatory diseases but does not have
anti-inflammatory properties. It is associated with increased cytokine activity, not suppression.

B. IL-6: IL-6 is a pro-inflammatory cytokine that plays a role in promoting immune responses and
inflammation. While it has some regulatory roles, it generally acts as a mediator of inflammation,
rather than being anti-inflammatory like IL-10.

C. TNF-alpha: TNF-alpha is another pro-inflammatory cytokine involved in inflammation and

immune responses. It is linked to the pathogenesis of several inflammatory diseases (such as
rheumatoid arthritis) and contributes to cytokine-mediated lethality in sepsis. It does not have
anti-inflammatory properties like IL-10.

D. IL-10 is the correct answer because it downregulates inflammation, suppresses the harmful
effects of cytokines like TNF-alpha and IL-1, and helps prevent excessive immune responses
that lead to autoimmune diseases and cytokine-mediated damage.

35. Which of the ff cytokines stimulate B cells to divide and differentiate into Ig-secreting cells?

a. IL-1
 c. IL-3

b. IL-2

d. IL-4

ANSWER:

A. IL-1: IL-1 is a pro-inflammatory cytokine that plays a role in immune response and
inflammation but does not directly stimulate B cell differentiation into antibody-secreting cells.

B. IL-2: IL-2 primarily promotes the proliferation of T cells, not B cells.

C. IL-3: IL-3 is involved in hematopoiesis and stimulates the growth and differentiation of
multiple blood cell types but does not specifically stimulate B cells to differentiate into
antibody-secreting cells.

D. IL-4 is the correct answer because it directly stimulates B cell activation and differentiation
into Ig-secreting plasma cells.

36. Which of the ff stimulates growth and differentiation of early lymphoid cells?
a. IL-2
c. IL-6
b. IL-3
d. IL-7

ANSWER:
A. IL-2: IL-2 primarily stimulates the proliferation and activation of T cells, not early lymphoid cell
differentiation.

B. IL-3: IL-3 stimulates hematopoiesis and the growth of multiple blood cell types, but it is not
specific to the differentiation of early lymphoid cells.

C. IL-6: IL-6 is involved in immune responses and inflammation but does not directly stimulate
the growth and differentiation of early lymphoid cells.

D. IL-7 is the correct answer because it specifically stimulates the growth and differentiation of
early lymphoid cells in both the bone marrow and thymus.

37. All are characteristics of anamnestic immune response, except:

a. rapid increase in IgM antibody levels
[Link] lag phase needed before antibody production
 c. results from repeated antigen exposure
d. also known as memory or booster response

ANSWER:
A. rapid increase in IgM antibody levels is incorrect because the anamnestic response is
characterized by a rapid increase in IgG antibodies, not IgM.

B. shorter lag phase needed before antibody production: In anamnestic responses, there is a
shorter lag phase because the immune system has memory from prior exposure, so it can
respond faster.

C. results from repeated antigen exposure: The anamnestic response is specifically a result of
prior exposure to the same antigen, which induces memory cell activation.

D. also known as memory or booster response: The anamnestic response is also called the
memory response because it involves memory cells that allow for a faster and stronger reaction
to a previously encountered pathogen.

38. Delayed-type hypersensitivity (DTH) skin tests are commonly used as measures of:
a. B cell function
b. T cell function
c. monocyte/marophage chemotaxis
d. IgE quantitation

ANSWER:
A. B cell function: B cells are primarily involved in antibody production, not in the T cell-mediated
DTH response. DTH is specifically a T cell response.

B. T cell function is correct because DTH responses are a measure of T cell-mediated immunity,
involving T cell activation and macrophage involvement.

C. monocyte/macrophage chemotaxis: While macrophages are involved in the inflammatory

response in DTH, the test itself is not designed to measure chemotaxis (the movement of
immune cells toward a chemical signal). It assesses T cell activation and response.

D. IgE quantitation: IgE is involved in allergic reactions, specifically type I hypersensitivity, and is
not relevant to the DTH response, which is mediated by T cells, not antibodies.

39. Post-streptococcal glomerulonephritis is classified as what type of hypersensitivity?

a. I
b. II
c. III
 d. IV

ANSWER:
A. Type I: Type I hypersensitivity involves IgE-mediated reactions, such as allergic reactions
(e.g., asthma, anaphylaxis). PSGN is not mediated by IgE or related to allergic responses.

B. Type II: Involves antibody-mediated cell destruction (e.g., hemolytic anemia), not immune
complex deposition like in PSGN.

C. Type III is correct because Post-streptococcal glomerulonephritis is caused by the deposition

of immune complexes in the kidneys, characteristic of type III hypersensitivity reactions.

D. Type IV: Is T cell-mediated, involving delayed reactions (e.g., contact dermatitis), not immune
complexes.

40. Which in vitro test measures IgE levels against a specific allergen?

a.​ Histamine release assay

b.​ Radioimmunosorbent test
c.​ Radioallergosorbent test
d.​ DTH skin test

(a) Histamine release assay (Incorrect): This measures histamine release from basophils
but does not directly quantify IgE levels.

(b) Radioimmunosorbent test (Incorrect): This is used to measure total IgE levels but not
allergen-specific IgE.

(c) Radioallergosorbent test (RAST) (Correct): RAST detects allergen-specific IgE in patient
serum, making it useful for allergy testing.

(d) DTH skin test (Incorrect): This measures cell-mediated immunity (T-cell response), not
IgE-mediated allergic reactions.

41. Which can be done to detect the coating of red cells by antibody as occurs in Type II
hypersensitivity-HTR?

a.​ IAT
b.​ DAT
c.​ Complement Fixation
d.​ ABO and Rh typing

(a) IAT (Incorrect): The Indirect Antiglobulin Test (IAT) detects free antibodies in serum,
but does not assess antibodies already bound to RBCs.
(b) DAT (Correct): The Direct Antiglobulin Test (DAT) detects antibodies or complement
coating RBCs, which occurs in Type II hypersensitivity reactions, such as Hemolytic
Transfusion Reactions (HTR).

(c) Complement Fixation (Incorrect): This tests for complement activation in serum but does
not assess antibody binding to RBCs.

(d) ABO and Rh typing (Incorrect): Blood typing determines antigen presence on RBCs but
does not detect antibody-mediated RBC destruction.

42. A patient deficient in the C3 complement component would be expected to mount a normal:

a.​ Type I and IV hypersensitivity response

b.​ Type Il and IV hypersensitivity response
c.​ Type I and III hypersensitivity response
d.​ Type Il and III hypersensitivity response

(a) Type I and IV (Correct):

●​ Type I (Immediate hypersensitivity) relies on IgE, mast cells, and histamine—not

complement.
●​ Type IV (Delayed-type hypersensitivity) is T-cell mediated and does not require
complement.

(b) Type II and IV (Incorrect):

●​ Type II (Cytotoxic hypersensitivity) involves complement activation for cell lysis,

which is impaired in C3 deficiency.
●​ Type IV remains normal, but Type II is affected.

(c) Type I and III (Incorrect):

●​ Type I remains normal, but Type III (Immune complex hypersensitivity) relies on C3
for immune complex clearance, making it abnormal in C3 deficiency.

(d) Type II and III (Incorrect):

●​ Both Type II and Type III require complement activation, so they would be impaired
in C3 deficiency.

43. How many mL of NSS is needed to be added to 0.2 mL serum to obtain 1:10 dilution?

a.​ 0.8 mL
 b.​ 1 mL
c.​ 1.8 mL
d.​ 2 mL

(a) 0.8 mL (Incorrect):

●​ If 0.8 mL NSS is added, the total volume would be 0.2 mL + 0.8 mL = 1.0 mL, making it
a 1:5 dilution, not 1:10.

(b) 1 mL (Incorrect):

●​ If 1 mL NSS is added, the total volume would be 0.2 mL + 1 mL = 1.2 mL, which does
not give a 1:10 dilution.

(c) 1.8 mL (Correct):

●​ This results in a total volume of 2.0 mL, achieving the correct 1:10 dilution.

(d) 2 mL (Incorrect):

●​ If 2 mL NSS is added, the total volume would be 0.2 mL + 2.0 mL = 2.2 mL, leading to a
1:11 dilution, which is too dilute.

44. Which of the ff are possible reasons when a patient tested negative on a test done on the
early course of infection?

1.​ Prozone phenomenon

2.​ Postzone phenomenon
3.​ Patient is immunocompromised
4.​ Patient has taken already antipyretic medications

a.​ 1 and 3
b.​ 2 and 3
c.​ 1 and 4
d.​ 2 and 4

(a) 1 and 3 (Incorrect): While an immunocompromised patient (3) may have a delayed or
absent immune response leading to a false-negative test, the prozone phenomenon (1) occurs
due to excessive antibodies interfering with antigen binding, which is more common in later
stages rather than early infection.
(b) 2 and 3 (Correct): The postzone phenomenon (2) occurs when there is an excess of
antigen over antibodies, preventing proper immune complex formation, and
immunocompromised patients (3) may not mount a detectable immune response, both
leading to false-negative results in early infection.

(c) 1 and 4 (Incorrect): Prozone phenomenon (1) is more relevant in later stages when
antibodies are in excess, and antipyretic medication (4) reduces fever but does not directly
affect antigen-antibody interactions, making this combination incorrect.

(d) 2 and 4 (Incorrect): Postzone phenomenon (2) is a valid cause of false negatives, but
antipyretic medication (4)only alleviates symptoms and does not interfere with serological test
results.

45. Which of the following scenarios shows prozone phenomenon in a serial dilution for Ab titer?

a.​ Tubes 1-5 show stronger reaction, while tubes 6-10 shows weaker reaction
b.​ Tubes 1-5 show weaker reaction, while tubes 6-10 shows stronger reaction
c.​ All tubes show strong reaction
d.​ All tubes show weak reaction

Tubes 1-5 show stronger reaction, while tubes 6-10 show weaker reaction (Incorrect):

●​ This pattern is typical of a normal serial dilution, where higher concentrations (lower
dilutions) yield stronger reactions, and as the sample is diluted further, the reaction
weakens.

Tubes 1-5 show weaker reaction, while tubes 6-10 show stronger reaction (Correct):

●​ This pattern represents the prozone phenomenon, where extremely high antibody
concentrations in the early tubes inhibit proper lattice formation, resulting in a weak
reaction, which improves upon dilution as the excess antibody is reduced.

All tubes show strong reaction (Incorrect):

●​ Uniform strong reactions across all tubes indicate that no prozone effect is occurring; the
antibody levels are optimal or consistently high across dilutions.

All tubes show weak reaction (Incorrect):

●​ Uniform weak reactions suggest that the antibody concentration is low throughout the
dilutions, not indicative of the prozone phenomenon.

46. In a two-fold dilution method, if tube 1 is undiluted, what will be the dilution on tube # 9?
 a.​ 1:64
b.​ 1:128
c.​ 1:256
d.​ 1:512

1:64 (Incorrect): This dilution corresponds to tube #7, not tube #9.

1:128 (Incorrect): This dilution is achieved in tube #8.

1:256 (Correct): In a two-fold dilution series starting with an undiluted tube, tube #9 is diluted
1:256.

1:512 (Incorrect): This dilution would be found in tube #10, one step beyond tube #9.

47. Compute for Clinical / Diagnostic Specificity of the test: TP: 130 TN: 45 FN: 20 FP: 5

a.​ 85%
b.​ 87%
c.​ 90%
d.​ 93%

85% (Incorrect): This value is lower than the calculated specificity.

87% (Incorrect): This value is lower than the calculated specificity.

90% (Correct): Matches the calculation using TN/(TN+FP) = 45/50.

93% (Incorrect): This value is higher than the calculated specificity.

48. Titer refers to the reciprocal of ____ dilution of serum which still produces a visible reaction
with a particular Ag

a.​ lowest
b.​ highest
c.​ equal
d.​ none of the above

Lowest (Incorrect): Titer is not based on the lowest dilution, but the highest dilution at which a
reaction is still visible.
Highest (Correct): Titer is defined as the reciprocal of the highest dilution of serum that still
produces a visible reaction with the antigen.

Equal (Incorrect): The term “equal dilution” is not used in titer determination.

None of the above (Incorrect): The correct description is provided by “highest.”

49. Antibody titer was taken twice from a patient with the results below. This is indicative of:

Acute phase: 8

Two weeks later: 128

a.​ Infection
b.​ Convalescence
c.​ Postzone phenomenon
d.​ Prozone phenomenon

a. Infection (Correct): A fourfold or greater increase in antibody titer (from 8 to 128) between
acute and convalescent phases strongly indicates an active or recent infection.

b. Convalescence (Incorrect): Convalescence refers to the recovery phase, where antibody

levels remain elevated but do not show a significant rise between two tests.

c. Postzone phenomenon (Incorrect): This occurs when excess antigen prevents proper
antibody binding, leading to false-negative results, but does not explain the rising titer.

d. Prozone phenomenon (Incorrect): This happens when excess antibodies interfere with
agglutination, causing weak or false-negative results, which does not apply here.

50. All of the following are similar when performing double immunodiffusion & single
immunodiffusion in petri plates, except

a.​ Antibodies are incorporated in agarose medium

b.​ Antigens and antibodies diffuse through agarose medium
c.​ Electric current is not applied and just rely on incubation
d.​ A visible precipitate is formed when Ag-Ab reactions occur

a. Antibodies are incorporated in agarose medium (Correct): In both single and double
immunodiffusion, antibodies and antigens are added to separate wells and diffuse toward each
other, rather than being pre-incorporated in the agarose.
b. Antigens and antibodies diffuse through agarose medium (True): Both methods rely on
passive diffusion of antigens and antibodies through the agarose gel.

c. Electric current is not applied and just rely on incubation (True): Unlike
electrophoresis-based techniques, these methods depend on passive diffusion and incubation
at appropriate conditions.

d. A visible precipitate is formed when Ag-Ab reactions occur (True): Both techniques
detect antigen-antibody interactions through the formation of a visible precipitin line.

51. Particle agglutination for qualitative detection of the capsular polysaccharide antigens of
Cryptococcus neoformans in CSF samples is an example of:

a.​ Agglutination inhibition

b.​ Coagglutination
c.​ Passive agglutination
d.​ Reverse passive agglutination

a. Agglutination inhibition (Incorrect): This method detects small antigens by preventing

visible agglutination rather than forming it, which does not apply to Cryptococcus antigen
testing.

b. Coagglutination (Incorrect): Coagglutination involves bacteria (e.g., Staphylococcus

aureus) coated with antibodies to detect antigens, but Cryptococcus antigen tests use latex
particles instead.

c. Passive agglutination (Incorrect): Passive agglutination involves particles coated with

antigens to detect antibodies, whereas Cryptococcus tests detect antigens.

d. Reverse passive agglutination (Correct): In this method, latex or other particles are coated
with antibodies to detect soluble antigens, such as Cryptococcus capsular polysaccharide in
CSF.

52. In countercurrent immunoelectrophoresis, the gel contains:

a.​ Antigen
b.​ Antibody
c.​ Both of the choices
d.​ None of the choices

Rationale:

a.​ Antigen is incorrect since it is not incorporated in the gel. Instead, it is put in a well and
moves towards the antibody during electrophoresis. If the antigen was pre-loaded in the
gel, it would be unable to move correctly for immunoprecipitation.
 b.​ In most CIE techniques, the gel is pre-loaded with an antibody, and the antigen is added
separately to the wells before electrophoresis. This setup allows for the fast transport of
antigens toward pre-embedded antibodies, resulting in precipitin lines in a short period of
time.
c.​ Both choices are incorrect since only the antibody is integrated into the gel. The antigen
is introduced individually and not pre-mixed with the gel matrix. If both were in the gel,
electrophoretic migration and appropriate antigen-antibody interaction would be
disturbed.
d.​ None of the choices because the gel does contain antibody, which play a crucial role in
the immunoprecipitation reaction during electrophoresis.

References:

Kurien, B. T., & Scofield, R. H. (2006). Western blotting and other protein detection
techniques. Methods in Molecular Biology, 536, 75-82. DOI: 10.1385/1-59745-007-3:75

Parikh, R. (2013). Immunoelectrophoresis techniques and their applications. Journal of

Clinical Laboratory Analysis, 27(3), 123-130.

53. In Ouchterlony diffusion, antibody is placed in:

a.​ Center well

b.​ Surrounding wells
c.​ Mixed with gel
d.​ Mixed with sample

Rationale:

a.​ In Ouchterlony diffusion, the antibody is put in the center well and the antigens in the
surrounding wells. As antigen and antibody diffuse, they meet at the zone of
equivalence, resulting in visible precipitin lines that show the existence of
antigen-antibody complexes. This approach enables the comparative study of several
antigens against a single antibody.
b.​ Surrounding wells are incorrect because they contain antigen, not antibody. The
antigens readily diffuse to the center, where they bind with the antibody.
c.​ Mixed with gel is incorrect because the antibody is not equally distributed throughout the
gel. Instead, it is put in a designated well to allow for regulated diffusion and contact with
antigens. Mixing the antibody into the gel would prevent clear precipitin lines from
forming.
d.​ Mixed with sample is incorrect since the antibody and antigen are put in separate wells
to facilitate diffusion towards each other. If they were pre-mixed, diffusion and precipitin
line formation would not work effectively.

References:
Ouchterlony, O. (1958). Diffusion-in-gel methods for immunological analysis. Progress in
Allergy, 5, 1-78.

Abul, K. A., & Lichtman, A. H. (2019). Basic Immunology: Functions and Disorders of the
Immune System. Elsevier.

54. Which of the following immunoassays does not require B/F separation?

a.​ Competitive EIA

b.​ Sandwich EIA
c.​ Indirect ELISA
d.​ EMIT

Rationale:

a.​ Competitive EIA is incorrect since competitive enzyme immunoassays require a

separation step to make distinctions between bound and free antigens before assessing
the signal.
b.​ Sandwich EIA is incorrect since sandwich ELISA involves washing procedures to
remove unbound analytes, making sure that only antigen-antibody complexes contribute
to the overall signal.
c.​ Indirect ELISA is incorrect since it also requires washing steps to eliminate binding
secondary antibodies, necessitating B/F separation.
d.​ EMIT is a homogeneous immunoassay, meaning that it does not require B/F separation.
Unlike heterogeneous immunoassays, which involve washing procedures to separate
bound from free analytes, EMIT depends on enzyme activity changes following antibody
binding. The enzyme-labeled antigen is active when unbound, but becomes blocked
when attached to an antibody. This enables for the direct measurement of enzyme
activity, eliminating the need for B/F separation.

References:

Tietz, N. W., & Rifai, N. (2017). Tietz Textbook of Clinical Chemistry and Molecular
Diagnostics. Elsevier.

Wild, D. (2013). The Immunoassay Handbook: Theory and Applications of Ligand Binding,
ELISA, and Related Techniques. Elsevier.

Wimsatt, J., & Kost, G. J. (2014). Immunoassays and Point-of-Care Testing. Clinical
Laboratory Medicine, 34(2), 363-386.

55. In ELISA, what component is used to be able to accurately quantify results?

a.​ Enzyme label

b.​ The solid phase
c.​ The second antibody
 d.​ The antigen

Rationale:

a.​ The enzyme label is the key component that enables precise measurement in ELISA.
The enzyme is attached to an antibody (or antigen in certain situations) and catalyzes a
reaction with a particular substrate, resulting in a detectable signal. The strength of this
signal is related to the concentration of target antigen or antibody in the sample. This
enzymatic process enables an accurate and sensitive detection of the analyte.
Horseradish peroxidase (HRP) and alkaline phosphatase (AP) are two often used
enzyme labels.
b.​ The solid phase is incorrect since the solid phase (typically a polystyrene microplate)
solely acts as a surface for antigen or antibody immobilization. It does not immediately
contribute to quantification.
c.​ The second antibody is incorrect because while a secondary antibody boosts specificity
and signal amplification, it does not directly provide the measurable signal. The enzyme
label coupled to the secondary antibody (in indirect or sandwich ELISA) is the
component responsible for quantification.
d.​ The antigen is incorrect because it is the target being measured. Its presence or
concentration is governed by the enzyme reaction rather than directly influencing the
observable signal.

References:

Lequin, R. M. (2013). Enzyme Immunoassay (EIA)/Enzyme-Linked Immunosorbent Assay

(ELISA). Clinical Chemistry, 59(4), 629-654.

Wild, D. (2013). The Immunoassay Handbook: Theory and Applications of Ligand Binding,
ELISA, and Related Techniques. Elsevier.

56. Which of the following is the reagent in direct immunofluorescence assay?

a.​ Known antigen conjugated with FITC

b.​ Known antibody conjugated with FITC
c.​ AHG conjugated with FITC
d.​ FITC only

Rationale:

a.​ Known antigen conjugated with FITC is incorrect because DFA detects antigens in a
sample using an antibody conjugated with FITC, not an antigen. Antigen labeling is more
common in competitive immunoassays, not direct immunofluorescence.
b.​ In direct immunofluorescence, a known antibody specific to the target antigen is
conjugated with FITC. This antibody binds directly to the antigen present in the sample.
Under a fluorescence microscope, the antigen-antibody complexes fluoresce, allowing
for visualization and identification of the target antigen.
 c.​ AHG coupled with FITC is incorrect since it is employed in the indirect
immunofluorescence test (IFA) rather than the direct immunofluorescence assay (DFA).
IFA uses a secondary antibody (such as AHG) coupled with FITC to detect a primary
antibody attached to the antigen.
d.​ D. FITC only is incorrect since FITC is a fluorophore and cannot bind to an antigen. It
must be coupled with an antibody to act in DFA.

References:

Goldstein, M. E., & Brown, R. D. (2014). Immunofluorescence techniques for antigen

detection. Methods in Molecular Biology, 1130, 157-172.

Bogdanovic, E., & Coons, A. H. (2013). Fluorescent antibody techniques in diagnostic

immunology. Clinical and Vaccine Immunology, 20(4), 473-478.

57. Indirect Immunofluorescence can be used to detect all of the following except:

a.​ Rickettsial antibodies

b.​ Treponemal antibodies
c.​ Surface markers in lymphocytes

Rationale:

a.​ Rickettsial antibodies is incorrect because IFA frequently identifies Rickettsial antibodies
in patient serum. The patient's antibodies (if present) attach to Rickettsial antigens on a
slide, which is then detected by a fluorescently labeled anti-human IgG or IgM.
b.​ Treponemal antibodies is incorrect because IFA is used to identify antibodies against the
bacteria Treponema pallidum, which causes syphilis. The Fluorescent Treponemal
Antibody-Absorption (FTA-ABS) test is an indirect immunofluorescence assay used for
syphilis detection.
c.​ Direct immunofluorescence or flow cytometry is the most effective method for detecting
lymphocyte surface markers, as fluorophore-conjugated monoclonal antibodies bind
directly to cell surface markers such as CD3, CD4, and CD8. Indirect
immunofluorescence is rarely used for this purpose since it needs a primary antibody,
which is not an efficient method for identifying cell surface markers.

References:

Paddock, C. D., & Childs, J. E. (2021). Rickettsial serology: Indirect immunofluorescence

assays. Clinical Microbiology Reviews, 34(2), e00023-20.

Fulwyler, M. J. (2015). Flow cytometry and fluorescent antibody techniques for

lymphocyte surface marker detection. Immunology Today, 18(3), 135-140.

58. Type of EIA which is used for the detection of antibodies to infectious agents (e.g., HIV,
HBV, HCV)
 a.​ Competitive
b.​ Indirect
c.​ Sandwhich
d.​ EMIT

Rationale:

a.​ Competitive is incorrect since competitive EIA is typically employed to identify tiny
compounds (e.g., hormones, drugs) rather than antibodies to infectious pathogens.
b.​ Indirect EIA is used to identify antibodies in a patient's serum. The process entails:
coating a solid phase (e.g., microplate well) with a known antigen (e.g., HIV, HBV, or
HCV antigens), adding the patient's serum if antibodies against the antigen are present,
they will bind to the coated antigen, adding an enzyme-labeled secondary antibody
(anti-human IgG or IgM conjugated with an enzyme, such as horseradish peroxidase
[HRP] or alkaline phosphatase), and substrate addition a colorimetric or
chemiluminescent reaction occurs, and the signal intensity correlates with the amount of
antibody present.
c.​ Sandwich is incorrect because sandwich EIA detects antigens rather than antibodies.
This approach needs two antibodies to attach to an antigen, which differs from the
format used to detect HIV, HBV, or HCV antibodies.
d.​ EMIT is incorrect since EMIT (Enzyme-Multiplied Immunoassay Technique) is a
homogeneous assay that detects tiny compounds like medicines without the need for a
separation step. It is not commonly used for identifying antibodies to infectious
pathogens.

Reference:

Wild, D. (2013). The Immunoassay Handbook: Theory and Applications of Ligand

Binding, ELISA, and Related Techniques. Elsevier.

59. In Competitive EIA, the enzyme activity is ______________ to antigen concentration.

a.​ Directly proportional

b.​ Indirectly proportional
c.​ Equivalent
d.​ Unrelated

Rationale:

a.​ Directly proportional is incorrect since the relationship in competitive EIA is inverse
rather than direct. A higher antigen concentration produces a lower signal.
b.​ When there is a larger quantity of antigen in the patient's sample, fewer enzyme-labeled
antigens may bind to the antibody, resulting in decreased enzyme activity and signal
strength. In contrast, when the antigen concentration is low, more tagged antigens bind,
resulting in increased enzyme activity and a greater signal.
 c.​ Equivalent is incorrect because enzyme activity does not equal antigen concentration.
Instead, it fluctuates in inverse proportion to it.
d.​ Unrelated is incorrect because enzyme activity is directly affected by antigen
concentration via competitive binding.

References:

Wild, D. (2013). The Immunoassay Handbook: Theory and Applications of Ligand Binding,
ELISA, and Related Techniques. Elsevier.

Tietz, N. W., & Rifai, N. (2017). Tietz Textbook of Clinical Chemistry and Molecular
Diagnostics. Elsevier.

60. What comprises the indicator system in an indirect ELISA for detecting antibody?

a.​ Enzyme-conjugated antibody + chromogenic substrate

b.​ Enzyme-conjugated antigen + chromogenic substrate
c.​ Enzyme + antigen
d.​ Substrate + antigen

Rationale:

a.​ Indirect ELISA aims to identify antibodies in a patient's serum. The indicator system
generates a quantifiable signal that corresponds to the presence and concentration of
the antibody. The standard indicator system in indirect ELISA is comprised of: (1) An
enzyme-conjugated secondary antibody that recognizes the patient's primary antibody.
(2) A chromogenic substrate that interacts with the enzyme, causing a color change that
is subsequently detected spectrophotometrically.
b.​ Enzyme-conjugated antigen + chromogenic substrate is incorrect since in indirect
ELISA, the enzyme label is attached to a secondary antibody rather than an antigen.
Antigen labeling is more prevalent in competitive ELISA.
c.​ Enzyme + antigen is incorrect since an enzyme cannot be detected without an antibody.
In addition, antigen is already immobilized on the plate in indirect ELISA.
d.​ Substrate + antigen is incorrect since the substrate does not interact directly with the
antigen. A discernible signal needs the use of an enzyme-conjugated antibody.

References:

Wild, D. (2013). The Immunoassay Handbook: Theory and Applications of Ligand Binding,
ELISA, and Related Techniques. Elsevier.

Tietz, N. W., & Rifai, N. (2017). Tietz Textbook of Clinical Chemistry and Molecular
Diagnostics. Elsevier.

61. What outcome results fro improper washing of a tube or well after adding the
enzyme-antibody conjugate in ELISA?
 a.​ Falsely decreased
b.​ Falsely increased
c.​ Result will be unaffected
d.​ Result is impossible to determine

Rationale:

a.​ Falsely decreased is incorrect because residual enzyme activity boosts the signal rather
than decreasing it.
b.​ Improper washing results in unbound enzyme-labeled antibodies. When the substrate is
introduced, these excess enzyme molecules catalyze the reaction, resulting in a larger
color change or signal that does not accurately represent analyte concentrations. This
results in a false positive or overestimation of the antibody or antigen under test.
c.​ Result will be unaffected is incorrect, because poor washing has an influence on the final
measurement by increasing background noise and non-specific binding.
d.​ Result is impossible to determine is incorrect because incorrect cleaning has a predicted
effect: erroneously raised readings.

References:

Wild, D. (2013). The Immunoassay Handbook: Theory and Applications of Ligand Binding,
ELISA, and Related Techniques. Elsevier.

Tietz, N. W., & Rifai, N. (2017). Tietz Textbook of Clinical Chemistry and Molecular
Diagnostics. Elsevier.

62. Autoantibody usually found in drug-induced lupus:

a.​ Anti-histone
b.​ Anti-dsDNA
c.​ Anti-phospholipid
d.​ anti-anti-SS-A/Ro

Rationale:

a.​ Anti-histone antibodies are present in 95% or more of drug-induced lupus cases.
b.​ Anti-dsDNA is incorrect because anti-dsDNA antibodies are primarily associated with
systemic lupus erythematosus (SLE) and are rarely found in drug-induced lupus. Their
presence in SLE is linked to renal involvement (lupus nephritis), which is uncommon in
drug-induced lupus.
c.​ C. Anti-phospholipid is incorrect because anti-phospholipid antibodies (such as
anticardiolipin and lupus anticoagulant) are more commonly associated with
antiphospholipid syndrome (APS) and SLE, not drug-induced lupus.
d.​ D. Anti-SS-A/Ro is incorrect because these antibodies are more commonly associated
with Sjogren’s syndrome and neonatal lupus, rather than drug-induced lupus.
References:

Solomon, L. A., & Sachs, B. (2015). Drug-induced lupus erythematosus: Incidence, clinical
manifestations, and management. Autoimmunity Reviews, 14(9), 872-879. DOI:
10.1016/[Link].2015.05.004

Burlingame, R. W., & Rubin, R. L. (2013). Autoantibodies to histones in drug-induced and

idiopathic lupus. Clinical & Experimental Immunology, 172(1), 1-7. DOI: 10.1111/cei.12043

63. Peripheral or rim pattern of immunofluorescence is characteristic of which ANA?

I.​ anti-dsDNA
II.​ Anti-RNP
III.​ Anti-DNP
IV.​ Anti-histone

a.​ 1 and 2
b.​ 1 and 3
c.​ 2 and 3
d.​ 2 and 4

RATIONALE:

a.​ 1 and 2 - Incorrect; while, anti-dsDNA does show a peripheral staining pattern,
Anti-RNP shows speckled pattern
b.​ 1 and 3 - both answers show peripheral and rim pattern
c.​ 2 and 3 - Incorrect pattern; while anti-DNP is a correct, Anti-RNP shows speckled
pattern
d.​ 2 and 4 - Incorrect pattern, Anti-RNP shows speckled pattern and Anti-Histone
shows homogeneous pattern

Reference: Tung, H. W., & Chen, M. F. (2017). Antinuclear antibody patterns and their
clinical significance. Journal of Clinical Laboratory Analysis, 31(5), e22073.
[Link]

64. FANA staining pattern if antibody against extractable nuclear antigens (ENA) is present:

a.​ Homogenous
b.​ Peripheral
c.​ Centromere
d.​ Speckled

RATIONALE:
 a.​ Homogenous - pattern does not occur when ENA is present; may occur in the presence
of ANA
b.​ Peripheral - pattern does not occur when ENA is present; may occur when Anti-dsDNA
antibodies are present
c.​ Centromere - does not occur when ENA is present; may occur in the presence of
anti-centromere antibodies (ACA)
d.​ Speckled - correct answer; occurs when ENA is present

65. Neonatal lupus, which occurs in up to 8% of babies born to pregnant women with SLE, is
associated with:

a.​ Anti-DNP
b.​ Anti-RNP
c.​ Anti-centromere
d.​ Anti-Ro (SS-A)

RATIONALE:

a.​ Anti-DNP - usually associated with systemic lupus erythematosus (SLE)

b.​ Anti-RNP - often associated with patients who have Mixed Connective Tissue Disease
(MCTD)
c.​ Anti-centromere - often associated with CREST syndrome
d.​ Anti-Ro (SS-A) - correct answer; often associated with Neonatal Lupus.

REFERENCE: Van den Hoogen, F., Khanna, D., Fransen, J., Johnson, S. R., Baron, M.,
Tyndall, A., ... & Furst, D. E. (2013). Classification criteria for systemic sclerosis: An
American College of Rheumatology/European League Against Rheumatism collaborative
initiative. Arthritis & Rheumatism, 65(11), 2737-2747. [Link]

66. Rheumatoid Factor is an _ antibody that targets the Fc portion of _.

a.​ IgG; IgM

b.​ IgM; IgG
c.​ IgD; IgM
d.​ IgA; IgG

RATIONALE:

a.​ IgG; IgM - Rheumatoid Factor is an IgM antibody, not an IgG antibody, and targets IgG
antibodies, not IgM.
b.​ IgM; IgG - This is the correct classification and function of Rheumatoid Factor.
c.​ IgD; IgM - Rheumatoid Factor is an IgM antibody, not an IgD antibody, and targets IgG
antibodies, not IgM.
d.​ IgA; IgG - Rheumatoid Factor is an IgM antibody, not an IgA antibody.
67. Affected tissue if there is high titer of anti-microsomal antibodies

a.​ Joints
b.​ Pancreas
c.​ Liver
d.​ Thyroid

RATIONALE:

a.​ Joints - ANA or Anti-nuclear antibodies will be present in high titers, not anti-microsomal
antibodies
b.​ Pancreas - If the pancreas is affected, anti-glutamic acid decarboxylase antibodies will
be present, and not anti-microsomal antibodies
c.​ Liver - ASMA or Anti-smooth muscle antibodies are present in high titers if the liver is the
affected tissue, and not anti-microsomal antibodies
d.​ Thyroid - Observed in conditions such as Hashimoto’s disease, Anti-microsomal
antibodies are usually present in high titers when the thyroid is affected.

REFERENCE: Fitzgerald, M., & Weinstein, M. (2023). Harrison's Principles of Internal

Medicine (21st ed.). McGraw-Hill Education.

68. What autoimmune condition is associated with this FANA staining pattern

-​ ---------
-​ ---------

a.​ Rheumatoid arthritis

b.​ CREST syndrome
c.​ Sjogrens syndrome
d.​ Neonatal lupus

RATIONALE:

a.​ Rheumatoid arthritis - the FANA staining pattern for rheumatoid arthritis is speckled and
homogenous
b.​ CREST syndrome - correct answer; centromere pattern matches the characteristics of
FANA for CREST syndrome
c.​ Sjogrens Syndrome - a fine-speckled pattern will be observed when a specimen positive
for Sjogren’s Syndrome is tested with FANA stain
d.​ Neonatal Lupus - a fine-coarse-speckled pattern will be observed when a specimen
positive for Neonatal Lupus is tested with FANA stain

69. Common variable immunodeficiency is characterized by absence of

 a.​ IgG and IgM
b.​ IgG and IgA
c.​ IgM and IgA
d.​ IgA only

RATIONALE:

a.​ IgG and IgM - IgM is not markedly decreased or absent in common variable
immunodeficiency (CVID)
b.​ IgG and IgA - Correct. Both antibodies are characteristically absent in CVID cases
c.​ IgM and IgA - Only IgA is absent in cases of CVID
d.​ IgA only - Incorrect, IgG is also included in the antibodies absent in CVID

Bonilla, F. A., & Ochs, H. D. (2017). Common variable immunodeficiency: Clinical and
immunologic features. Journal of Allergy and Clinical Immunology, 139(3), 1111-1121.
[Link]

70. All of the following tests can evaluate phagocytic disorders except

a.​ DHR assay

b.​ NBT dye
c.​ Mitogen stimulation
d.​ CD18 assay

RATIONALE:

a.​ DHR assay - is able to evaluate phagocytic disorders

b.​ NBT dye - can evaluate phagocytic disorders
c.​ Mitogen stimulation - is not known or applied when evaluating phagocytic disorders as it
primarily evaluates T-cell function, not phagocytes
d.​ CD18 assay - can evaluate phagocytic disorders

Reference: Flores, A. L., & Nelson, R. P. (2007). Evaluation of immune function: T-cell and
phagocytic assays. Journal of Clinical Immunology, 27(4), 345-351.
[Link]

71. The BinaxNOW Strep A Test immunochromatographic assay is used for detection of S.
pyogenes _ from _ specimens

a.​ Antigen; serum

b.​ Antigen; throat swab
c.​ Antigen; saliva
d.​ Antibody; serum

RATIONALE:
 a.​ Antigen; serum - Serum specimens are not the first choice for Strep A diagnosis
b.​ Antigen; throat swab - both answers are correct and accurate to the use of BinaxNOW
Strep A test
c.​ Antigen; saliva - salivary specimens are not the first choice for Strep A diagnosis
d.​ Antibody; serum - S. pyogenes antibodies are not being detected when detecting a Strep
infection

REFERENCE: Abbott. (2020). BinaxNOW™ Strep A: Product insert. Abbott Laboratories.

[Link]

72. This method measures antibodies against five extracellular streptococcal antigens;
anti-streptolysin (ASO), anti-hyaluronidase (AHase), anti-streptokinase (ASKase),
anti-nicotinamide-adenine nucleotide (anti-NAD), and anti-DNAse B antibodies.

a.​ Lateral flow assay

b.​ Streptozyme
c.​ ASO-hemolytic test
d.​ BinaxNOW

RATIONALE:

a.​ Lateral flow assay - this method is generally a broader group of test which also includes
Streptozyme—which is more specific for ASO, ASKase, anti-NAD, and anti-DNAse
antibodies
b.​ Streptozyme - the Streptozyme test is a serological assay that can measure antibodies
against several streptococcal antigens, but it is more commonly used to detect
antibodies to five specific streptococcal antigens
c.​ ASO-hemolytic test - this test specifically measure ASO antibodies only, and not the rest
of the antibodies involved
d.​ BinaxNOW - the BinaxNOW Strep A test is designed to detect the presence of
streptococcal antigens, not antibodies

REFERENCE: Patel, R., & Ziegler, S. (2020). Evaluation of rapid diagnostic tests for
streptococcal infections. Clinical Infectious Diseases, 71(9), 1187-1195.
[Link]

73. The gold standard for the serological diagnosis of Rocky Mountain Spotted Fever

a.​ Culture
b.​ PCR
c.​ IFA
d.​ Weil-Felix Test

RATIONALE:
 a.​ Culture - this is not a serological method of diagnosis nor is it used for diagnosing
Rocky Mountain Spotted Fever
b.​ PCR - this is a molecular diagnostic technique instead of a serological technique
c.​ IFA - the Indirect Immunofluorescence Assay is the gold standard for detection
and confirmation of Rocky Mountain Spotted Fever
d.​ Weil-Felix Test - not a standard test used to detect nor confirm Rocky Mountain
Spotted Fever

REFERENCE: Turgeon, M. L. (2017). Immunology and serology in laboratory medicine (5th

ed.). Elsevier.

74. In Weil-Felix Test, the rickettsial antibodies are able to cross-react with:

a.​ Proteus antigens

b.​ Streptococcal antigens
c.​ Rickettsial antigens
d.​ All of the choices

REFERENCE:

a.​ Proteus antigens - Proteus vulgaris OX19 antigen is the most commonly associated
antigen with cross-reaction in the Weil-Felix test
b.​ Streptococcal antibodies - not the most common antigen detected in Weil-Felix Test
c.​ Rickettsial antigens - although detected in WeilFelix Test, this is not as common as cross
reacting with Proteus antigens
d.​ All of the choices - incorrect, only A is correct.

75. Used as confirmatory test for samples that initially test positive for Lyme disease by EIA or
IFA:

A. Nucleic acid amplification - primarily used for confirming Lyme disease in synovial fluid or
tissue in cases of Lyme arthritis but not as a routine confirmatory test following a positive
EIA/IFA.

B. Culture - impractical for routine clinical use and is not used as a confirmatory test after a
positive EIA/IFA.

C. Western Blot - Western blot is a serological test used to detect antibodies against specific
Borrelia burgdorferi proteins. It serves as the confirmatory test after an initial positive result from
an enzyme immunoassay (EIA) or indirect immunofluorescence assay (IFA). A positive Western
blot involves detecting IgM or IgG antibodies at specific bands that correlate with Lyme disease.

D. RIA - not part of the standard diagnostic protocol for Lyme disease.
Reference: Centers for Disease Control and Prevention. (2022). Lyme disease: Two-step
laboratory testing process.

76. Reference: Radolf, J. D., Deka, R. K., Anand, A., & Sadzewicz, L. (2021). Treponema
pallidum: Genomic insights and implications for clinical microbiology. Clinical Microbiology
Reviews, 34(2), e00146-20.

77. Volume of serum placed in an 18-mm circle on plastic-coated disposable card for RPR
testing

A. 0.010 mL - Too small a volume for adequate antigen-antibody interaction, leading to

unreliable results.

B. 0.05 mL - ensures a uniform spread of the sample within the 18-mm circle, allowing the
antigen to interact effectively with antibodies if present.

C. 0.10 mL - Double the recommended volume, which may cause excessive spreading of the
sample, potentially leading to non-uniform reactions.

D. 0.5 mL - Far too large a volume, which would flood the test circle and invalidate the test.

Reference: Larsen, S. A., Steiner, B. M., & Rudolph, A. H. (1995). Laboratory diagnosis and
interpretation of tests for syphilis. Clinical Microbiology Reviews, 8(1), 1–21.

78. Reagin antibodies are non-specific _____________ antibodies directed against cardiolipin
constituent of treponemal lipids but usually derived from its host

A. IgG - While IgG is involved, it is not the only antibody involved. IgM also plays a role,
especially in the early stages.

B. IgE - IgE is associated with allergic reactions and parasitic infections, not with syphilis or
cardiolipin-directed responses.

C. IgG or IgE - IgE is not relevant to syphilis, making this combination incorrect.

D. IgG or IgM - Both IgG and IgM are involved in the immune response to lipid antigens such as
cardiolipin.

Reference: Turgeon, M. L. (2018). Clinical hematology and fundamentals of hemostasis (6th

ed.). Wolters Kluwer.
79. Most specific test for diagnosis of neurosyphilis using CSF as sample:

A. RPR - non-treponemal test similar to VDRL but is less sensitive and specific when used on
CSF.

B. VDRL - A positive VDRL in CSF is considered diagnostic of neurosyphilis, although the

sensitivity is moderate, meaning a negative result does not rule out the disease.

C. EIA - detects treponemal antibodies but is not used to diagnose neurosyphilis using CSF.

D. PCR - can detect T. pallidum DNA in CSF but has limited sensitivity and is not routinely
recommended for neurosyphilis diagnosis.

Reference: Turgeon, M. L. (2018). Clinical hematology and fundamentals of hemostasis (6th

ed.). Wolters Kluwer.

80. Which syphilis testing requires inactivation of serum samples?

A. RPR and VDRL - RPR does not require inactivation due to choline chloride, which neutralizes
complement.

B. VDRL and FTA-ABS - require serum inactivation to prevent complement interference and
ensure accurate results.

C. EIA and CIA - Do not involve complement, so inactivation is unnecessary.

D. FTA-ABS and TP-PA - TP-PA an agglutination assay that does not require inactivation.

Reference: World Health Organization. (2016). Laboratory diagnosis of sexually transmitted

infections, including human immunodeficiency virus.

81. Which tests are included in Reverse Sequencing Testing Algorithm for syphilis?

1.​ VDRL
2.​ RPR
3.​ EIA
4.​ TP-PA

A. 1 and 2 - RPR is non-treponemal test used in the reverse algorithm, but VDRL is not included
in this algorithm.
B. 2 and 3 - This option is incomplete because it excludes TP-PA, which is used to confirm
discordant results.

C. 1, 2 and 3 - Correctly included but this option misses TP-PA, which is necessary for
confirming discordant results.

D. 2, 3 and 4 - RPR is Non-treponemal test to detect active syphilis. EIA is Treponemal test
used for initial screening. And TP-PA is Confirmatory test to resolve discordant results. This
option correctly includes all the tests used in the reverse sequence algorithm.

Reference: Centers for Disease Control and Prevention. (2021). Sexually transmitted diseases
treatment guidelines. U.S. Department of Health and Human Services.

82. What is the use of Nichol's strain of treponeme used in FTA-ABS?

A. acts as sorbent - Reiter strain (non-pathogenic treponeme) is used as the sorbent to remove
cross-reacting antibodies.

B. acts the reagent antigen - In the FTA-ABS test, Nichol’s strain is fixed onto a microscope
slide to detect antibodies (IgG and IgM) in the patient’s serum.

C. use to attach FITC - is used to label the anti-human globulin, which attaches to bound
antibodies, but the Nichol’s strain does not perform this function.

D. use to destroy native complement - Serum inactivation (56°C for 30 minutes) is done to
destroy complement, not the role of the Nichol’s strain.

Reference: Turgeon, M. L. (2018). Linne & Ringsrud’s clinical laboratory science: The basics
and routine techniques (7th ed.). Elsevier.

83. Composition of the antigen complex used in VDRL and RPR:

1.​ Cardiolipin
2.​ Cholesterol
3.​ Lecithin
4.​ Charcoal

A. 1 and 2 - This option is incomplete because it excludes lecithin, which is essential for
enhancing specificity.

B. 1 and 3 - Cholesterol is missing, which is necessary for stabilizing the antigen complex.
C. 1, 2 and 3 - The antigen complex in VDRL and RPR consists of cardiolipin, cholesterol, and
lecithin, which together detect reagin antibodies in syphilis.

D. 1, 2, 3 and 4 - Charcoal is only included in RPR, not VDRL. VDRL uses a microscopic
reading method without charcoal.

Reference: Turgeon, M. L. (2018). Linne & Ringsrud’s clinical laboratory science: The basics
and routine techniques (7th ed.). Elsevier.

84. Positive result observed in T. pallidum particle agglutination (TP-PA) test:

A. latticelike structure that spreads (smooth mat) - A positive TP-PA test shows a lattice-like
structure that spreads as a smooth mat, indicating agglutination of antibody-coated particles.

B. gel particle settle to the bottom of the well (compact button) - A compact button indicates a
negative result, where no agglutination occurs, and the particles settle at the bottom.

C. macroscopic flocculation - Flocculation occurs in non-treponemal tests such as RPR and

VDRL, where reagin antibodies react with cardiolipin [Link]-PA does not involve
flocculation.

D. microscopic fluorescence - Microscopic fluorescence is observed in FTA-ABS (Fluorescent

Treponemal Antibody-Absorption test), not TP-PA.

Reference: American Society for Microbiology (ASM). (2020). Manual of clinical microbiology
(12th ed.). Carroll, K. C., et al.

85. Presence of Anti-HBc IgM & undetectable HBsAg during the window phase indicates
_______ infection

A. Acute / Recent - This is a transient phase where HBsAg becomes undetectable while
anti-HBc IgM remains the primary marker of infection. Anti-HBc IgM is produced in the early
phase of infection, indicating a recent or acute HBV infection.

B. Chronic - Anti-HBc IgG predominates in chronic infections, not anti-HBc IgM. Chronic
infection is also associated with the persistence of HBsAg and possibly HBeAg.

C. Past - A past infection is characterized by the presence of anti-HBc IgG and anti-HBs, not
anti-HBc IgM.

D. Latent - Latent HBV infection is defined by the presence of HBV DNA in the liver without
detectable HBsAg or clinical symptoms, not by the presence of anti-HBc IgM.
Reference: Centers for Disease Control and Prevention (CDC). (2021). Hepatitis B: Serologic
testing and interpretation.

86. First antibody detected in HBV infection is directed against:

A. HBsAg - surface antigen and is not the target of the first antibody response; instead,
antibodies to HBsAg (anti-HBs) appear later during recovery.

B. HBeAg - secreted protein associated with viral replication but is not the initial antigen that
elicits the early antibody response.

C. HBcAg - is the target of the first antibody produced in HBV infection; although HBcAg itself is
not detectable in serum, the immune response rapidly produces anti-HBc IgM, making it the
earliest serological marker in the infection.

D. HBV DNA - genetic material of the virus and does not induce an antibody response, as
antibodies are directed against protein antigens.

Reference: Turgeon, M. L. (2018). Linne & Ringsrud’s Clinical Laboratory Science: The Basics
and Routine Techniques (7th ed.). Elsevier.

87. First serologic evidence of convalescent phase of HBV infection is indicated by presence of:

a.​ Anti-HBc IgM

b.​ Anti-HBc IgG
c.​ Anti-HBe
d.​ Anti-HBs

Answer:

a.​ It appears early in acute infection and is a marker of recent HBV infection. It declines as
the infection resolves and is not the first marker of the convalescent phase.
b.​ It replaces IgM over time and persists for life, whether the patient recovers or develops
chronic infection. it is not specific for the convalescent phase, as it remains detectable
even after recovery.
c.​ It appears as HBeAg disappears, marking the transition from active infection to the
recovery phase, making it the first serologic evidence of the convalescent phase.
d.​ It is the last marker to appear and signals full recovery and immunity. However, Anti-hBe
appears first in the convalescent phase, making it Anti-HBs a late indicator rather than
the earliest sign.

Reference:
 ●​ Dienstag, J. L. (2008). Hepatitis B virus infection. New England Journal of Medicine,
359(14), 1486-1500.

88. Which of the following viruses is rarely implicated in transfusion-associated hepatitis?

a.​ Hepatitis A
b.​ Hepatitis B
c.​ Hepatitis C
d.​ None of the choices

Answer:

a.​ Hepatitis A is transmitted primarily through the fecal-oral route, not through blood
transfusion. It is an acute, self-limiting infection and does not lead to chronic hepatitis,
making it rarely associated with transfusion-related hepatitis.
b.​ HBV is a major cause of transfusion-associated hepatitis due to its presence in blood
and bodily fluids. Despite screening, transfusion-related HBV cases still occur, especially
in occult infections.
c.​ Before routine screening of blood donations, HCV was the leading cause of
transfusion-associated hepatitis. It is transmitted via contaminated blood products, and
although screening has drastically reduced cases, it remains a concern in high-risk
settings.
d.​ HAV is rarely implicated in transfusion-associated hepatitis due to its transmission route
being primarily through ingestion of contaminated food or water rather than blood
transfusion.

Reference :

●​ Hollinger, F. B., & Emerson, S. U. (2007). Hepatitis A virus. Clinical Microbiology

Reviews, 20(3), 361–385.

89. Anti-HBs can be detected in which cases?

1. Previous vaccination for HBV. 3. Past infection of HBV

2. Previous injection of HB Ig. 4. Low-level carriers of HBV

a. 1 and 2. c. 1, 2 and 3

b. 1 and 3. d. 1, 2, 3 and 4

Answer:

a.​ Anti-HBs is the main antibody produced in response to HBV vaccination. The vaccine
contains Hepatitis B surface antigen (HBsAg), stimulating the immune system to develop
protective antibodies (Anti-HBs). Hepatitis B immune globulin (HBIG) is administered for
 post-exposure prophylaxis. It contains preformed Anti-HBs antibodies, which provide
short-term passive immunity but do not indicate an actual immune response.
b.​ Anti-HBs is the main antibody produced in response to HBV vaccination. The vaccine
contains Hepatitis B surface antigen (HBsAg), stimulating the immune system to develop
protective antibodies (Anti-HBs). Individuals who have recovered from HBV infection
naturally develop Anti-HBs along with Anti-HBc IgG. This indicates immunity due to past
exposure.
c.​ Anti-HBs is the main antibody produced in response to HBV vaccination. The vaccine
contains Hepatitis B surface antigen (HBsAg), stimulating the immune system to develop
protective antibodies (Anti-HBs). Hepatitis B immune globulin (HBIG) is administered for
post-exposure prophylaxis. It contains preformed Anti-HBs antibodies, which provide
short-term passive immunity but do not indicate an actual immune response. Individuals
who have recovered from HBV infection naturally develop Anti-HBs along with Anti-HBc
IgG. This indicates immunity due to past exposure.
d.​ 1,2 and 3 correct and 4 is Incorrect because chronic low-level carriers, HBsAg is
persistently present, which typically prevents the production of Anti-HBs. The presence
of both markers is uncommon but can occur in rare cases with immune fluctuations.

Reference :

●​ Raimondo, G., Pollicino, T., Romano, L., & Zanetti, A. R. (2010). A 2010 update on
occult Hepatitis B infection. Pathologie Biologie, 58(4), 254–260.

90. Evaluates the effectiveness of antiviral therapy in patients with chronic hepatitis B:

a. HBsAg. c. liver enzyme measurement

b. HBeAg. [Link] load testing

answer:

a.​ HBsAg is used to diagnose active HBV infection but is not the best marker for evaluating
the effectiveness of antiviral therapy. Persistent HBsAg positivity indicates chronic
infection, while its loss (HBsAg seroclearance) is a sign of recovery, but this occurs late
in treatment.
b.​ HBeAg is a marker of HBV replication and infectivity. Its loss (HBeAg seroconversion)
indicates reduced viral replication and is a treatment goal in HBeAg-positive patients.
However, antiviral therapy effectiveness is best monitored through viral load testing.
c.​ ALT (alanine aminotransferase) and AST (aspartate aminotransferase) levels help
assess liver inflammation and damage. While decreasing ALT levels may indicate
treatment response, they do not directly measure viral suppression, making them a
secondary marker.
d.​ HBV DNA viral load testing directly measures the amount of HBV in the blood, making it
the most reliable method for evaluating antiviral therapy effectiveness. A decline in HBV
DNA levels indicates a successful response to treatment.
reference :

●​ Liaw, Y. F., & Chu, C. M. (2009). Hepatitis B virus infection. The Lancet, 373(9663),
582-592.

91. Heterophile antibodies associated with infectious mononucleosis are _____ antibodies
produced as a result of polyclonal B-cell activation and reacts with horse, sheep and bovine red
blood cell.

a.​ IgM
b.​ IgG
c.​ IgA
d.​ IgE

answer:

a.​ Heterophile antibodies associated with infectious mononucleosis (IM) are IgM
antibodies. They are produced due to Epstein-Barr virus (EBV)-induced polyclonal B-cell
activation and react with horse, sheep, and bovine red blood cells (RBCs) in the classic
Paul-Bunnell test. The Monospot test, commonly used for IM diagnosis, detects these
heterophile IgM antibodies.
b.​ IgG antibodies are produced later in EBV infections and are associated with long-term
immunity rather than acute heterophile responses. IgG antibodies against viral capsid
antigen (VCA-IgG) appear later and persist for life, helping differentiate past from recent
infections.
c.​ IgA antibodies are primarily found in mucosal secretions (e.g., saliva, tears) and play a
role in mucosal immunity. They are not typically involved in the heterophile antibody
response in infectious mononucleosis.
d.​ IgE antibodies are involved in allergic reactions and parasitic infections, not in the
immune response to EBV or heterophile antibody production. There is no significant
association between IgE and infectious mononucleosis.

reference :

●​ Hoagland, R. J. (1960). The heterophile antibody test in infectious mononucleosis.

Journal of the American Medical Association, 174(6), 546-549.

92. Receptor utilized by EBV to infect lymphocytes:

a.​ CCR5
b.​ CXCR4
c.​ CD21
d.​ both A and B

answer:
 a.​ CCR5 (C-C chemokine receptor 5) is a co-receptor primarily used by HIV-1 to enter
CD4+ T cells, not Epstein-Barr virus (EBV). It plays a role in HIV infection and immune
cell trafficking, but it is not involved in EBV entry.
b.​ CXCR4 (C-X-C chemokine receptor 4) is another HIV co-receptor that allows T-tropic
strains of HIV to infect CD4+ T cells. Although important in immune cell signaling,
CXCR4 is not the receptor for EBV.
c.​ CD21 is the primary receptor utilized by Epstein-Barr virus (EBV) to infect B
lymphocytes. EBV binds to CD21, which is also the C3d complement receptor, allowing
viral entry and subsequent infection. This receptor is highly expressed on B cells, making
them the main target of EBV.
d.​ CCR5 and CXCR4 are HIV co-receptors and do not play a role in EBV infection. Since
CD21 is the correct receptor, this option is incorrect.

reference:

●​ Fingeroth, J. D., Weis, J. J., Tedder, T. F., Strominger, J. L., Biro, P. A., & Fearon, D. T.
(1984). Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2.
Proceedings of the National Academy of Sciences, 81(14), 4510-4514.

93. Serologic marker for convalescence in infectious mononucleosis:

a.​ Anti-VCA IgM

b.​ Anti-ENA
c.​ ANTI-EBNA IgG
d.​ Heterophile antibodies

answer:

a.​ Anti-VCA IgM appears early in infectious mononucleosis (IM) and is a marker of acute
infection, not convalescence. It typically declines within 4–6 weeks after infection.
b.​ Anti-ENA antibodies are associated with autoimmune diseases like systemic lupus
erythematosus (SLE) and Sjogren’s syndrome, not EBV infection or infectious
mononucleosis.
c.​ Anti-EBNA IgG develops late in infection (typically 3–6 months after onset) and persists
for life, making it a key serologic marker of convalescence and past infection in infectious
mononucleosis.
d.​ Heterophile antibodies (detected by the Monospot test) appear early in infection and
decline within weeks to months. They are a diagnostic marker for acute IM but not for
convalescence.

reference:

●​ Luzuriaga, K., & Sullivan, J. L. (2010). Infectious mononucleosis. New England Journal
of Medicine, 362(21), 1993-2000.
94. Which of the following results are observed in Davidsohn differential test in case of EBV
infection?

a.​ heterophile titer is low after absorption with guinea pig kidney cells
b.​ heterophile titer is high after absorption with guinea pig kidney cells
c.​ heterophile titer is high after absorption with beef erythrocytes
d.​ heterophile titer is high after absorption with both guinea pig kidney cells and beef
erythrocytes

answer:

a.​ In the Davidsohn Differential Test, heterophile antibodies from infectious mononucleosis
(IM) are NOT absorbed by guinea pig kidney cells. This means that after absorption, the
heterophile titer remains high in EBV infections. A low titer would indicate a different type
of heterophile antibody, such as Forssman antibodies, which are absorbed by guinea pig
kidney cells.
b.​ In EBV infection, heterophile antibodies are not absorbed by guinea pig kidney cells but
are absorbed by beef erythrocytes. This means the heterophile titer remains high after
absorption with guinea pig kidney cells, confirming infectious mononucleosis. The test
helps differentiate IM from Forssman antibodies, which would be absorbed by guinea pig
kidney cells, leading to a lower titer.
c.​ Beef erythrocytes absorb heterophile antibodies associated with infectious
mononucleosis. If the titer remains high after absorption with beef erythrocytes, it
suggests a different type of heterophile antibody, not one specific to EBV.
d.​ If both guinea pig kidney cells and beef erythrocytes fail to absorb the heterophile
antibodies, this indicates another type of heterophile reaction (e.g., Forssman or serum
sickness antibodies). In infectious mononucleosis, the titer should be high after guinea
pig kidney cell absorption but low after beef erythrocyte absorption.

reference :

●​ Hoagland, R. J. (1960). The heterophile antibody test in infectious mononucleosis.

JAMA, 174(6), 546-549. [Link]

95. All of the following tests can detect heterophile antibodies associated with IM, except:

a.​ Paul-Bunnel
b.​ Davidsohn differential
c.​ Monospot
d.​ Viral load test

answer:

a.​ The Paul-Bunnell test is a classic heterophile antibody test used for infectious
mononucleosis (IM). It detects IgM heterophile antibodies that agglutinate sheep RBCs
 in patients with EBV infection. This test is not highly specific, as Forssman and serum
sickness antibodies may also cause agglutination.
b.​ This test is a modification of the Paul-Bunnell test to differentiate heterophile antibodies
caused by IM from those due to Forssman antibodies or serum sickness. IM-associated
heterophile antibodies react with beef erythrocytes but not guinea pig kidney cells,
confirming EBV infection.
c.​ The Monospot test is a rapid slide agglutination test for heterophile antibodies in IM. It
detects heterophile antibodies using horse RBCs, offering faster and more specific
results than the Paul-Bunnell test. This test is widely used in clinical settings for
diagnosing IM.
d.​ Viral load tests measure EBV DNA in blood using polymerase chain reaction (PCR). This
test is used for quantifying viral replication, especially in cases of chronic EBV infection,
post-transplant lymphoproliferative disorders, or EBV-associated malignancies. It does
not detect heterophile antibodies, making it the correct answer to this question.

reference :

●​ Gulley, M. L., & Tang, W. (2008). Using Epstein-Barr viral load assays to diagnose,
monitor, and prevent posttransplant lymphoproliferative disorder. Clinical Microbiology
Reviews, 21(2), 263-267. [Link]

96. _____ forms the numerous knobs or spikes that protrude from the outer envelope, whereas
______ is a transmembrane glycoprotein that spans the inner and outer membrane of HIV.

a.​ Gp41;Gp120
b.​ Gp120;Gp41
c.​ p24;p17
d.​ p17;p24

answer:

a.​ Gp41 is the transmembrane glycoprotein, but it does not form the external knobs or
spikes. Instead, it plays a role in membrane fusion during viral entry. The correct
sequence should have Gp120 first, as it is responsible for forming the external spikes.
b.​ Gp120 is the external glycoprotein that forms the numerous knobs or spikes on the outer
envelope of HIV. It binds to CD4 receptors on host cells to initiate infection. Gp41 is a
transmembrane glycoprotein that spans the viral membrane and is essential for fusion
with the host cell membrane.
c.​ p24 is the capsid protein of HIV, forming the viral core rather than being part of the
envelope. p17 is the matrix protein found just beneath the viral envelope, helping
maintain structural integrity.
d.​ p17 is the matrix protein, and p24 is the capsid protein, both of which are found inside
the virus. Neither of them forms the external spikes or spans the viral membrane.

reference :
 ●​ Kwong, P. D., Wyatt, R., Robinson, J., Sweet, R. W., Sodroski, J., & Hendrickson, W. A.
(1998). Structure of an HIV gp120 envelope glycoprotein in complex with the CD4
receptor and a neutralizing human antibody. Nature, 393(6686), 648-659.

97. The first antibodies to be detected in HIV infection are directed against ____, followed by
antibodies to _____.

a.​ Gp41;p24
b.​ Gp120;p24
c.​ p55;gp41
d.​ p17;gp120

answer:

a.​ The first antibodies to be detected in HIV infection are primarily directed against Gp41,
the transmembrane glycoprotein involved in viral fusion. Antibodies to p24 (the capsid
protein) appear shortly after but tend to decline as the infection progresses due to
immune complex formation.
b.​ Gp120-specific antibodies develop later in infection, as Gp120 is highly variable and
subject to immune evasion mechanisms. p24 antibodies appear before Gp120
antibodies, making this option incorrect.
c.​ p55 is the Gag polyprotein precursor that gets cleaved into p24 (capsid), p17 (matrix),
and p7/p6 (nucleocapsid proteins). Antibodies against p55 are not among the first to
appear in HIV infection.
d.​ p17 (matrix protein) is part of the viral core and does not elicit the earliest detectable
antibodies in HIV infection. Gp120 antibodies develop later than Gp41 antibodies due to
its high mutation rate and immune evasion strategies.

reference :

●​ Fiebig, E. W., Wright, D. J., Rawal, B. D., Garrett, P. E., Schumacher, R. T., Peddada, L.,
& Busch, M. P. (2003). Dynamics of HIV viremia and antibody seroconversion in plasma
donors: Implications for diagnosis and staging of primary HIV infection. AIDS, 17(13),
1871-1879.

98. In HIV testing using Western blot, which of the following is detected in patient serum?

a.​ HIV antigens

b.​ HIV antibodies
c.​ HIV nuclear proteins
d.​ Enzyme Reverse transcriptase

answer:

a.​ The Western blot test for HIV detects antibodies, not antigens. HIV antigens, such as
p24, are detected in earlier-stage tests like ELISA or p24 antigen assays, but not in
 Western blot. Western blot works by identifying patient antibodies that bind to specific
HIV proteins transferred onto a membrane.
b.​ Western blot is a confirmatory test for HIV that detects antibodies against HIV proteins in
patient serum. When HIV is present, the immune system produces antibodies targeting
viral proteins like gp41, gp120, p24, and p31. If antibodies bind to specific HIV proteins
on the Western blot strip, the test is considered positive.
c.​ HIV nuclear proteins, such as integrase or nucleocapsid proteins, play a role in HIV
replication and integration but are not the primary targets of Western blot testing.
Western blot focuses on HIV envelope (gp120, gp41) and core (p24) proteins, which
stimulate a strong antibody response in infected individuals.
d.​ Reverse transcriptase (RT) is an enzyme produced by HIV, but Western blot does not
detect it. RT activity is measured in laboratory assays to study HIV replication but is not
used for serological HIV diagnosis.

reference:

●​ Klatzmann, D., Barre-Sinoussi, F., & Montagnier, L. (1984). The human T-lymphotropic
retrovirus, LAV, in AIDS patients. Nature, 312(5992), 767-770.
[Link]

99. Which of the following can be used as confirmatory test for HIV?

a.​ Rapid assay with combined Ag/Ab detection of HIV-1 and HIV-2
b.​ Rapid assay to differentiate anti-HIV-1 and HIV-2
c.​ Indirect EIA to detect p24 antigen
d.​ both A and B

answer:

a.​ This test detects both HIV antigen (p24) and antibodies (anti-HIV-1 and anti-HIV-2) but is
used as an initial screening test, not a confirmatory test. It is part of fourth-generation
HIV testing, which improves early detection by identifying p24 antigen before antibodies
appear. However, a confirmatory test like Western blot or HIV-1/HIV-2 differentiation
assay is required after a positive result.
b.​ This differentiation assay is used as a confirmatory test after a positive HIV screening
test (e.g., ELISA or Ag/Ab combo assay). It distinguishes between HIV-1 and HIV-2
antibodies, which is important because HIV-2 progresses more slowly and requires
different management. The Geenius HIV-1/2 Confirmatory Assay is an example of this
type of test.
c.​ p24 antigen detection is used in early HIV diagnosis but is not a confirmatory test. While
it helps detect acute HIV infection, it does not distinguish between HIV-1 and HIV-2 and
does not confirm a reactive screening test. Confirmatory testing requires antibody
differentiation or Western blot.
 d.​ While option B (HIV-1/2 differentiation assay) is a confirmatory test, option A (Ag/Ab
detection) is only a screening test. Since option A does not confirm HIV infection, this
answer is incorrect.

reference :

●​ Pandori, M. W., Hackett, J., Louie, B., et al. (2009). Detection of HIV using p24 antigen
testing. Journal of Clinical Virology, 46(1), 26-31.

100. Prime targets of HIV:

a.​ Tc cells
b.​ Th cells
c.​ B cells
d.​ NK cells

answer:

a.​ Cytotoxic T cells (CD8+ T cells) play a crucial role in killing virus-infected cells, including
those infected with HIV. However, HIV does not primarily target CD8+ T cells because
they lack the CD4 receptor, which is necessary for viral entry.
b.​ Helper T cells (CD4+ T cells) are the primary targets of HIV because the virus uses the
CD4 receptor, along with CCR5 or CXCR4 co-receptors, to enter and infect these cells.
HIV infection leads to a progressive decline in CD4+ T cells, weakening the immune
system and eventually causing AIDS.
c.​ B cells are responsible for antibody production, but they are not directly infected by HIV.
However, HIV indirectly affects B cells by disrupting CD4+ T cell function, leading to poor
antibody responses and increased susceptibility to infections.
d.​ Natural killer (NK) cells are part of the innate immune system and help eliminate infected
cells. They are not a primary target of HIV because they do not express CD4, but HIV
can impair NK cell function over time, reducing their effectiveness.

reference :

●​ Douek, D. C., Roederer, M., & Koup, R. A. (2009). Emerging concepts in the
immunopathogenesis of AIDS. Annual Review of Medicine, 60, 471-484.
 BLOODBANK POST TEST RATIONALE (MARCH 29,
2025)
1. Additive solutions (SAGM) should be added to _ within 72 hrs of collection

A.​ Whole blood

B.​ RBCs
C.​ Plasma
D.​ Platelet concentrate

ANSWER:

A.​ SAGM (Saline-Adenine-Glucose-Mannitol) is not added directly to whole blood,

but rather to red blood cell (RBC) components after separation.
B.​ SAGM is an additive solution specifically formulated for storage of packed
red blood cells.
C.​ Plasma is typically frozen (e.g., fresh frozen plasma or FFP) and does not
require additive solutions like SAGM.
D.​ Platelets are stored at 20–24°C with constant agitation, and are not preserved
with SAGM.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

2. Which of the following blood preservatives contains adenine to maintain ATP levels?

A.​ ACD
B.​ CPD
C.​ CP2D
D.​ Rejuvenation solution

ANSWER:

A.​ It does not contain adenine, so it does not support prolonged ATP synthesis in
red cells.
B.​ They may contain adenine, but they are not standard preservatives and are
used after storage, not for initial preservation.
C.​ CP2D alone is not a direct source of adenine.
D.​ Adenine supports ATP synthesis, which is crucial for maintaining RBC
membrane integrity and viability during storage.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

3. Storage temperature needed for Deglycerolized RBCs

 A.​ 0°C
B.​ -18°C
C.​ -65°C
D.​ 1-6°C

ANSWER:

A.​ 0°C is not an acceptable storage temperature for any blood component under
standard blood banking practices.
B.​ This is the minimum temperature for frozen plasma (e.g., fresh frozen
plasma).
C.​ This temperature is used for long-term storage of frozen RBCs with
high-glycerol cryopreservation.
D.​ This temperature range helps preserve red cell viability and function without
the risk of hemolysis.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

4. All of the following blood Ac-preservative offers a shelf-life of 21 days except:

A.​ ACD
B.​ CPD
C.​ CP2D
D.​ Heparin

ANSWER:

A.​ It supports red blood cell metabolism by providing glucose for ATP production.
B.​ Like ACD, it allows for a 21-day shelf life of stored blood at 1–6°C.
C.​ Like CPD, it offers a 21-day shelf life, unless combined with an additive solution
to extend storage further.
D.​ It does not preserve red cell metabolism and therefore does not allow for
storage beyond a few hours.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

5. All are examples of biochemical changes that RBCs undergo during refrigerated storage
except for:

A.​ loss of ATP and glucose

B.​ b. loss of RBC deformability
C.​ accumulation of lactic acid and a decrease in pH of blood
D.​ accumulation of 2,3-DPG that can impair oxygen release to tissues

ANSWER:
 A.​ Adenosine triphosphate (ATP) is essential for maintaining red blood cell (RBC)
membrane integrity and ion transport.
B.​ RBCs need to be flexible to pass through small capillaries.
C.​ RBCs rely on anaerobic glycolysis, which produces lactic acid as a byproduct.
D.​ 2,3-Diphosphoglycerate (2,3-DPG) levels decrease, not accumulate.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

6. How many pairs of autosomes do humans have?

A.​ 20
B.​ 21
C.​ 22
D.​ 23

ANSWER:

A.​ Humans have more than 20 pairs of autosomes.

B.​ 21 pairs of autosomes is still below the correct number.
C.​ Humans have 22 pairs of autosomes, which are non-sex chromosomes
D.​ 23 is the total number of chromosome pairs, not all autosomes.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

7. All of these blood group genes are located on autosomes, except:

A.​ Duffy
B.​ Kidd
C.​ YT
D.​ XG

ANSWER:

A.​ The Duffy blood group is encoded by the FY gene, which is located on
chromosome 1 (an autosome).
B.​ The Kidd blood group system is governed by the SLC14A1 gene, located on
chromosome 18 (an autosome).
C.​ The YT blood group is associated with the ACHE gene, found on chromosome
7 (an autosome).
D.​ The XG blood group is encoded by the XG gene, which is located on the X
chromosome, not an autosome.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

8. McLeod phenotype which demonstrates weak Kell antigens, is inherited by which manner?
 A.​ Autosomal dominant
B.​ Autosomal recessive
C.​ Autosomal codominant
D.​ X-linked recessive

ANSWER:

A.​ McLeod phenotype is not inherited this way and does not involve autosomes.
B.​ The McLeod phenotype is not autosomal, but X-linked, and affects mostly
males due to their single X chromosome.
C.​ The Kell blood group system, while codominant, is not responsible for
McLeod, which involves a separate gene.
D.​ The McLeod phenotype results from mutations or deletions in the XK gene on
the X chromosome.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

9. All are correct about RBC phenotypes, except:

A.​ describes what antigens are present on RBCs

B.​ tested by using monoclonal reagent antibodies
C.​ expression is based from inheritance of blood group genes
D.​ Examples: AO, BO, AB, 0O, DCe/dce

ANSWER:

A.​ The RBC phenotype refers to the specific antigens that are present on the
surface of red blood cells (RBCs).
B.​ RBC phenotyping is typically performed using monoclonal reagent antibodies
that are specific for different blood group antigens.
C.​ The phenotype of an individual’s red blood cells is determined by the inheritance
of specific blood group genes.
D.​ The correct notation for ABO phenotypes should be AO, BO, AB, OO, where
the second letter represents the allele inherited from the other parent.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

10. Immune A and B alloantibodies differ from naturally occurring A and B alloantibodies in that
immune antibodies:

A.​ Are generally IgG rather than IgM

B.​ Are unable to cross the placenta
C.​ Can be enhanced in reactivity by incubation at 4C
D.​ Causes direct agglutination at room temperature
ANSWER:

A.​ Immune alloantibodies are typically IgG antibodies produced in response to

exposure to foreign antigens, such as in transfusion or pregnancy.
B.​ The majority of immune alloantibodies (IgG) can cross the placenta and may
cause hemolytic disease of the fetus and newborn (HDFN) if they target antigens
on fetal red blood cells.
C.​ Immune alloantibodies, particularly those of the IgG type, are more likely to be
enhanced in reactivity when incubated at lower temperatures, such as 4°C.
D.​ Immune antibodies, specifically those of the IgG class, are typically unable to
agglutinate red blood cells directly at room temperature unless conditions such
as enzyme treatment or other specific conditions are applied.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

11. Which of the following has the potential to cause production of alloantibodies?

A.​ Pregnancy
B.​ Recent blood donation
C.​ Penicillin drug binding to RBC membrane
D.​ Paroxysmal Cold hemoglobinuria

ANSWER:

A.​ Pregnancy can lead to the production of alloantibodies because of potential

exposure to fetal red blood cells, especially during childbirth or in cases of
placental bleeding.
B.​ A recent blood donation can lead to the production of alloantibodies if the donor's
blood has antigens that the recipient does not possess.
C.​ Penicillin binding to the red blood cell (RBC) membrane can create a
drug-induced immune response.
D.​ Paroxysmal Cold Hemoglobinuria is a rare condition in which exposure to cold
temperatures leads to the formation of autoantibodies, typically directed against
the P antigen on red blood cells.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

12. Clinically significant antibodies react at and has potential to cause HTR or HDN.

A.​ 4-22C
B.​ 4-30C
C.​ 22-30C
D.​ 30-37C
ANSWER:

A.​ Antibodies that react at temperatures between 4°C and 22°C are typically of the
IgM class.
B.​ Antibodies that react within this temperature range can include IgG antibodies
such as those involved in Rh incompatibility (e.g., anti-D).
C.​ Antibodies reacting in the range of 22°C to 30°C can also be IgG antibodies, but
these are typically associated with more specific antigen-antibody interactions.
D.​ Antibodies that react between 30°C and 37°C are IgG antibodies, which are
typically the most clinically significant in transfusion medicine.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

13. Most naturally occurring antibodies are IgM cold agglutinins and they could be hemolytic
when active at

A.​ 4C
B.​ 22C
C.​ 37C
D.​ 56C

ANSWER:

A.​ Naturally occurring IgM cold agglutinins are most active at 4°C.
B.​ At 22°C, which is closer to room temperature, naturally occurring IgM cold
agglutinins are still active and can cause agglutination of red blood cells.
C.​ At 37°C, naturally occurring IgM cold agglutinins are usually not active
because IgM antibodies primarily act at lower temperatures (below body
temperature).
D.​ At 56°C, naturally occurring IgM cold agglutinins would be inactivated due to
the denaturation of the antibody structure at this temperature.

Reference: Modern blood banking and transfusion practices by Harmening (7th ed)

14. Reaction pattern / Principle observed in Gel technology:

a.​ Hemolysis
b.​ Hemagglutination
c.​ RBC Pellet
d.​ Immune adherence

RATIONALE

A. Hemolysis – Incorrect. Hemolysis is a destructive process, not the principle of gel testing.
B. Hemagglutination – Correct. Gel technology is based on hemagglutination where
agglutinated red cells are trapped in the gel matrix.

C. RBC Pellet – Incorrect. A pellet indicates a negative result, not the principle.

D. Immune adherence – Incorrect. This principle is associated with complement fixation, not gel
testing.​
Reference: Harmening, D. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.,
Chapter 6.

15. All are advantages of Gel technology except for:

a.​ No need for washing of RBCs before AHG phase

b.​ No need to use antisera and reagent red cells
c.​ can be used in automated instruments
d.​ results are stable for 2-3 days

RATIONALE:

A. No need for washing of RBCs before AHG phase – True. Washing steps are eliminated in gel
testing.

B. No need to use antisera and reagent red cells – Incorrect. Gel technology still requires
antisera and reagent RBCs for antigen-antibody reactions.

C. Can be used in automated instruments – True. Gel technology is compatible with automation.

D. Results are stable for 2–3 days – True. Gel cards can be stored and re-evaluated.​
Reference: AABB Technical Manual, 20th ed., Chapter 21.

16. Gel technology: Presence of a layer of agglutinated RBCs at the top of the gel and RBC
pellet at the bottom of microtube

a.​ 0
b.​ 1+
c.​ 2+
d.​ MF

RATIONALE

A. 0 – Incorrect. All RBCs form a pellet at the bottom (no agglutination).

B. 1+ – Incorrect. Shows few agglutinates dispersed in the lower part of the gel.
C. 2+ – Incorrect. Agglutinates are in the middle of the gel column.

D. MF – Correct. Mixed field shows both agglutinated cells (at the top) and unagglutinated cells
(pellet at bottom), typical in transfused or chimeric patients.​
Reference: Harmening, D., Modern Blood Banking, 7th ed.

17. Which is incorrect about postzone phenomenou?

a.​ excess in antigen in the test system

b.​ leads to false-negative reactions
c.​ remedy is by performing serum dilution
d.​ caused by the use of too concentrated RCS

RATIONALE:

A. Excess in antigen in the test system – Correct (true statement). The postzone phenomenon
occurs when there is an excess of antigen, preventing adequate cross-linking with antibodies
and leading to a false-negative result.

B. Leads to false-negative reactions – Correct (true statement). The imbalance in

antigen-antibody ratio can inhibit visible agglutination even when antibodies are present.

C. Remedy is by performing serum dilution – Incorrect (answer). Serum dilution is typically used
to correct a prozone effect (excess antibody), not postzone. For postzone, delaying testing to
allow antigen clearance or increasing antibody concentration may be more appropriate.

D. Caused by the use of too concentrated RBCs – Correct (true statement). This might sound
like prozone, but in the context of postzone, it can also refer to high cell concentrations acting as
excess antigen surfaces.

Reference: Harmening, D. (2019). Modern Blood Banking, 7th ed.

18. Which of the following can show antigen dosage?

a.​ Jka
b.​ MNSs
c.​ Fya
d.​ D
a.​ 1 and 2
b.​ 1 and 3
c.​ 1 , 2 and 3
d.​ 1, 2, 3 and 4
RATIONALE:

a. Jk (Kidd system) – Correct. Kidd antigens can show dosage, though they are sometimes
weakly expressed.

b. MNSs – Correct. M and N, as well as S and s antigens, exhibit dosage effects.

c. Fya (Duffy system) – Correct. Fya shows stronger reactions with homozygous cells.

d. D (Rh system) – Incorrect. The D antigen generally does not exhibit dosage.

Reference: Harmening, D. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.,
Chapter 7 – Blood Group Serology.

19. Anti-M antibodies react optimally on which RBC phenotype?

a.​ M+N+
b.​ M+N-
c.​ M-N+
d.​ Mk

RATIONALE:

a. M+N+ – Incorrect. This heterozygous expression shows weaker reactivity than homozygous
cells.

b. M+N- – Correct. Anti-M reacts more strongly with homozygous M+ cells due to dosage.

c. M−N+ – Incorrect. These cells do not express the M antigen.

d. Mk – Incorrect. Mk phenotype lacks both M and N antigens.

Reference: AABB Technical Manual, 20th ed., Table on antigen expression and antibody
reactivity.

20. Based on the following antigram, which cell is heterozygous for M?

a.​ Cell 1
b.​ Cell 2
c.​ Cell 3
d.​ None of the above

RATIONALE:
a. Cell 1 is correct. A heterozygous expression (M+N+) usually shows weaker agglutination with
anti-M than a homozygous M+N− cell.

b. Cell 2 is incorrect. If Cell 1 shows a weaker reaction with anti-M compared to another cell that
is M+N−, this suggests it is heterozygous.

c. Cell 3 is incorrect. The principle here is antigen dosage—where antibodies react more
strongly with cells that are homozygous for the target antigen.

d. None of the above is incorrect because Cell 1 is considered as the correct answer based
from the given table.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

21. Which of the following antigens is not enhanced not destroyed by the use of enzymes?

a.​ Xga
b.​ Fya
c.​ K
d.​ I

RATIONALE:

a. Xga – Usually destroyed by enzymes like papain or ficin.

b. Fya – Duffy antigens are destroyed by enzymes.

c. K (Kell) – Not enhanced nor destroyed by enzyme treatment; enzyme treatment has no
significant effect.

d. I – Enhanced by enzymes.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

22. Dyes commonly used in AHG:

1.​ Patent blue

2.​ Tartrazine
3.​ Acriflavine
4.​ Methyl green
a.​ 1 and 2
b.​ 1 and 3
c.​ 2 and 3
 d.​ 4 only

RATIONALE:

A. 1 and 2. Patent blue and Tartrazine are dyes used in some AHG reagents, but they are less
common compared to others. Incorrect choice as it doesn't fully match the typical dyes used in
AHG.

B. I and 3 Patent blue is used as a colorant, but Acriflavine is also used in some AHG reagents.
Incorrect as Patent blue is not a standard dye used in AHG reagents.

C. 2 and 3. Tartrazine (a yellow dye) and Acriflavine (fluorescent yellow) are commonly used in
AHG reagents for distinguishing between different types of AHG reagents and for easier
detection in agglutination testing.

D. 4 only. Methyl green is not typically used in AHG reagents. The other dyes (like Tartrazine
and Acriflavine) are more common.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

23. Which is not used as enhancement medium or potentiator in BB procedures?

a.​ LISS
b.​ PEG
c.​ NSS
d.​ AHG

RATIONALE:

A. LISS is a widely used enhancement medium in blood banking, lowering ionic strength to help
antibodies come in closer contact with RBCs.

B. PEG is used to enhance antibody binding by concentrating antibodies and decreasing the
distance between RBCs. It enhances hemagglutination.

C. NSS is not an enhancement medium but is often used as a diluent or for washing cells. It
does not play the same role as LISS or PEG in potentiating reactions.

D. AHG is used after the RBCs have been sensitized with antibodies to detect agglutination. It is
not a potentiator. It detects the agglutination by binding to the antibody-coated RBCs.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.
24. Which is incorrect about the use of enhancement media?

a.​ Increases the chance of hemagglutination

b.​ increases the zeta potential between RBCs
c.​ Increases the degree of RBC sensitization
d.​ Decreases the needed incubation time of test mixture

RATIONALE:

A. Increases the chance of hemagglutination is incorrect. Enhancement media such as LISS

and PEG increase the chance of hemagglutination by facilitating antibody attachment to RBCs,
reducing the distance between them and increasing sensitivity.

B. Increases the zeta potential between RBCs is correct. Enhancement media decrease the
zeta potential, which is the negative charge between RBCs. This helps RBCs come closer
together and facilswitates agglutination. Increasing the zeta potential would make it harder for
agglutination to occur.

C. Increases the degree of RBC sensitization is incorrect. Enhancement media like LISS and
PEG help increase RBC sensitization by reducing the ionic strength, which allows antibodies to
bind more efficiently.

D. Decreases the needed incubation time of test mixture is incorrect. By reducing zeta potential,
enhancement media decrease the incubation time needed for agglutination because they allow
antibodies to react with RBCs more efficiently.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

25. In the traditional or tube method, LISS is added to the test mixture just before _______
phase.

a.​ immediate spin

b.​ 37C incubation
c.​ washing phase
d.​ antiglobulin

RATIONALE:

A. Immediate spin is incorrect. LISS is not added during the immediate spin phase. This phase
is used to centrifuge the test tube immediately after mixing. LISS is added before incubation, not
before the spin.
B. 37°C incubation is Correct. LISS (Low Ionic Strength Solution) is added just before the 37°C
incubation phase. LISS reduces the ionic strength of the solution, which allows for more efficient
antibody binding to the red cells, enhancing the antibody-antigen reaction.

C. Washing phase is Incorrect. The washing phase occurs after incubation, where excess
reagents and unbound antibodies are removed. LISS is not added during the washing phase.

D. Antiglobulin is Incorrect. The antiglobulin phase (AHG phase) occurs after incubation and
washing to detect sensitization by antibodies or complement. LISS is added before incubation,
not during the antiglobulin phase.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

26. AHG has specificity for the _______ acting as a bridge & cross linking red cells

a.​ Fab portion of IgG

b.​ Fc portion of IgG
c.​ Fab portion of IgM
d.​ Fc portion of IgM

RATIONALE:

A. Fab portion of IgG is Incorrect. The Fab portion of IgG binds to antigens, but AHG does not
specifically act on this portion. The Fab region is responsible for antigen recognition, not for
cross-linking the red cells.

B. Fc portion of IgG is Correct. AHG (Anti-Human Globulin) binds specifically to the Fc portion of
IgG, which allows it to cross-link red cells that have been sensitized with antibodies. The Fc
portion is crucial for the agglutination process in the AHG phase.

C. Fab portion of IgM is Incorrect. AHG does not specifically act on the Fab portion of IgM. IgM
is generally large and pentameric, and AHG usually reacts with IgG antibodies and their Fc
region in blood bank testing.

D. Fc portion of IgM is Incorrect. While IgM has an Fc portion, AHG specifically targets IgG
antibodies and does not cross-link cells using IgM's Fc portion.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

27. What is the action of AHG reagent?

a.​ cross-links red cells that have become sensitized with antibody or complement.
 b.​ renders the red cell membrane more hydrophobic, allowing red cells to come closer
together.
c.​ reduces the zeta potential of the red cell, allowing closer approach of red cells.
d.​ release sialic acid from the red cell membrane decreasing negative chargers between
the cells

RATIONALE:

A. Correct. AHG (Anti-Human Globulin) reagent acts by cross-linking red cells that have been
sensitized with antibodies or complement. It binds to the Fc portion of IgG antibodies (or
complement) on the surface of red blood cells, resulting in agglutination (clumping of red blood
cells), which is crucial in detecting sensitized red blood cells in blood bank procedures.

B. Incorrect. AHG does not act by altering the hydrophobic nature of red blood cell membranes.
Its action is related to cross-linking sensitized cells, not modifying membrane properties to
increase proximity.

C. Incorrect. While reducing the zeta potential can enhance red blood cell agglutination, this is
typically achieved by using enhancement media like LISS or PEG, not AHG. AHG's primary
action is cross-linking sensitized red blood cells rather than modifying the zeta potential.

D. Incorrect. AHG does not release sialic acid from the red blood cell membrane. The sialic acid
on red blood cells contributes to their negative charge, but AHG's role is to bind to antibodies or
complement on sensitized cells, leading to agglutination.

Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

28. Anti-Pl present in patient serum is usually detected in which reaction phase?
a.​ Immediate spin
b.​ 37C
c.​ AHG
d.​ Both B and C

ANSWER:
A.​ Immediate spin is the correct answer because Anti-Pl is typically an IgM antibody,
which reacts best at colder temperatures (like room temperature), leading to
agglutination in the immediate spin phase.
B.​ This phase is typically used to detect IgG antibodies. IgG antibodies are usually
warm-reactive and would not be best detected at this temperature if the antibody
is IgM, like Anti-Pl. Therefore, the detection of Anti-Pl would not be optimal at
37°C.
C.​ This phase is used to detect IgG antibodies that have already formed
antigen-antibody complexes but have not caused agglutination due to the small
 size of IgG antibodies. Since Anti-Pl is usually an IgM antibody, the AHG phase
would not be the most appropriate for detecting it.
D.​ Since the best phase for detecting Anti-Pl is the immediate spin phase, neither
37°C nor the AHG phase is optimal for this purpose, making this option incorrect.

29. Polyspecific AHG cannot be used in:

a. detecting Hemolytic disease of the newborn
b. detecting Hemolytic transfusion reaction
c. confirming in vivo coating of red cells
d. differentiation of cell surface coating (complement or IgG)

ANSWER:
A.​ Polyspecific AHG can be used in the detection of HDN, as this condition is often
associated with IgG antibodies (such as anti-Rh antibodies) coating the red blood
cells of the newborn. Polyspecific AHG can detect the presence of these IgG
antibodies on the RBC surface.
B.​ Polyspecific AHG is also used in detecting hemolytic transfusion reactions,
which often involve the in vivo coating of red blood cells with IgG antibodies
from the transfusion. Therefore, polyspecific AHG can be used to detect this
phenomenon.
C.​ Polyspecific AHG is used to confirm in vivo coating of red blood cells, such as
in conditions like autoimmune hemolytic anemia (AIHA) or transfusion reactions.
It can detect both IgG antibodies and complement on the RBC surface, making
it useful for this purpose.
D.​ differentiation of cell surface coating (complement or IgG) is the correct
answer because polyspecific AHG cannot distinguish between complement and
IgG coating on red blood cells. It reacts with both, making it unsuitable for this
specific differentiation.

30. At the end of an antiglobulin test, IgG-coated control cells are added to the negative tests
and centrifuged. If no agglutination occurs, this means that:
a. test is valid
b. inhibition by autoantibodies
c. AHG reagent was neutralized
d. Enzyme treatment destroyed the reaction

ANSWER:
A.​ If no agglutination occurs when adding IgG-coated control cells, it suggests a
problem with the test. A valid test should show agglutination of the IgG-coated
control cells. Therefore, the absence of agglutination indicates an issue, not that
the test is valid.
 B.​ Autoantibodies typically interfere with the patient's own red blood cells and
can cause false positive or negative results, but they wouldn't generally cause an
issue with the IgG-coated control cells. Autoantibodies may cause problems in
interpreting the test results, but they are unlikely to be the reason for the lack of
agglutination with the control cells.
C.​ If no agglutination occurs after adding IgG-coated control cells, it likely
means the AHG reagent was neutralized, which is why c is correct. The
absence of agglutination indicates that the AHG reagent failed to bind to the red
blood cells, usually due to neutralization or interference, making this the correct
interpretation.
D.​ Enzyme treatment can alter or destroy red blood cell antigens, potentially
affecting antigen-antibody reactions, but it wouldn’t typically prevent AHG
reagent from working. The problem here is more likely to be neutralization of
the AHG reagent, not destruction of the antigen-antibody reaction caused by
enzyme treatment.

31. Specimen used in Direct Coomb's Test:

a. Patient red cells only
c. Patient red cells plus donor serum
b. Patient serum only
d. Patient serum plus donor red cells

ANSWER:
A.​ The Direct Coombs Test specifically uses patient red cells only (option a) to
detect whether there are any antibodies or complement already bound to those
cells. Therefore, a is the correct answer, and b, c, and d are incorrect because
they do not reflect the correct procedure or specimen for a Direct Coombs Test.
B.​ The Direct Coombs Test does not use patient serum because it is testing for
antibodies or complement already boundto the patient’s red blood cells. In
contrast, Indirect Coombs Test uses patient serum to detect free antibodies
in the serum, not bound to red blood cells.
C.​ This combination is not used in the Direct Coombs Test. This scenario more
closely resembles an Indirect Coombs Testwhere the donor serum would be
used to test for antibodies in the serum against the patient's red blood cells.
D.​ This combination is also more appropriate for an Indirect Coombs Test. In the
Indirect Coombs Test, patient serum is mixed with donor red cells to detect
antibodies in the serum that may react with antigens on the donor's red blood
cells. This is a test for free antibodies in the serum, not antibodies bound to the
patient's red blood cells.
32. Positive results of DAT are monitored by a panel using _____ anti-IgG and anti-C3d to
determine the specific type of protein sensitizing the cell.
[Link] AHG
b. Polyclonal AHG
c. Monospecific AHG
d. Polyspecific AHG

ANSWER:
A.​ Monoclonal AHG is derived from single clones of B cells and is highly specific for
detecting a single type of antibody or complement. However, the correct reagent
in this context is monospecific AHG, which is focused on detecting either IgG or
C3d, not both at once. Monoclonal reagents may not always be used in panels
that differentiate between IgG and complement.
B.​ Polyclonal AHG is a mixture of antibodies derived from different sources that
can recognize multiple epitopes on IgG or complement. While it can detect
general antibody or complement binding, it doesn't allow the specific
identification of IgG vs. complement. Therefore, it wouldn't be as useful for this
purpose when the goal is to differentiate between IgG and complement.
C.​ Monospecific AHG is used to specifically detect either IgG or complement
(C3d) on the red blood cells, allowing the lab to differentiate between the two.
This is why c. Monospecific AHG is the correct answer. Monoclonal,
polyclonal, and polyspecific AHG reagents do not provide the specificity
needed for this distinction.
D.​ Polyspecific AHG is a mixture of both anti-IgG and anti-complement
antibodies. While this reagent can be used to detect the presence of either IgG
or complement on red blood cells, it cannot specifically differentiate between
them. This is because polyspecific AHG reacts with both types of proteins
simultaneously. For identifying the exact type of sensitization (IgG vs.
complement), monospecific AHG is the correct choice.

33. Centrifugation time needed for antiglobulin test:

a. 20 seconds
b. 60 seconds
c. 5 minutes
d. 10-15 minutes

ANSWER:
A.​ The correct centrifugation time for the antiglobulin test is 20 seconds (option a),
as it is the ideal time needed to help the red blood cells settle without causing
over-compaction, which would obscure the detection of agglutination. The other
options (b, c, d) involve times that are either too long and not suitable for
accurate test results.
 B.​ 60 seconds is generally too long for the antiglobulin test. A longer centrifugation
time could lead to excessive pellet formation, making it difficult to assess the
agglutination reaction. Therefore, 20 seconds is the optimal time to ensure the
cells settle without overcompaction.
C.​ 5 minutes is much too long for the antiglobulin test. Prolonged centrifugation
might lead to improper interpretation of the results, as the cells would form too
tight of a pellet, obscuring any agglutination reactions. Additionally, this amount
of time is unnecessary for the test’s purpose.
D.​ 10-15 minutes is an even longer period and is far too much time for the
antiglobulin test. As with the 5-minute option, it would cause over-pelletization
and interfere with the test’s ability to properly detect agglutination.

34. Composition of Check cells / Coombs' control:

a. Type O red cells sensitized with IgG Anti-D .
b. Type O red cells sensitized with IgM Anti-D
c. Type AB red cells sensitized with IgG Anti-D
d. Type AB red cells sensitized with IgM Anti-D

ANSWER:
A.​ The correct composition of check cells (Coombs' control cells) is Type O red cells
sensitized with IgG anti-D (option a) because these cells are used to confirm the
proper function of the AHG reagent by detecting IgG sensitization. Type O cells
are used for consistency, and IgG anti-D is the appropriate antibody for this
control, as it reacts specifically with the AHG reagent. The other options (b, c, d)
are incorrect due to issues with antibody type or red blood cell type.
B.​ IgM antibodies are typically larger and can cause agglutination on their own at
room temperature. However, IgM anti-D is not the typical reagent used for
check cells in antiglobulin testing. The purpose of check cells is to verify the
action of IgG antibodies, as IgG is the more clinically relevant antibody for many
transfusion-related tests. Additionally, IgM anti-D would not be suitable for the
check cell process because IgM typically does not require AHG for agglutination
detection, as it agglutinates directly at room temperature.
C.​ Type AB red cells would not be used for check cells because AB red blood
cells have both A and B antigens, which could introduce variability and affect test
results, especially if the test is being done with other red blood cell samples that
are Type O. Type O red cells are preferred for consistency in Coombs' testing
because they lack A and B antigens.
D.​ Similar to option c, Type AB red cells are not ideal for check cells because they
could introduce antigenic variability. Additionally, IgM anti-D would cause
agglutination without the need for AHG, making it unsuitable for use in the control
process for verifying AHG reagent function.
35. ABO IgM antibodies are frequently hemolytic because of:
a. their dimeric structure
b. molecule's five antigen binding sites
c. Immune reactivity at 4-22C and 37C
d. their efficient ability to fix complement

ANSWER:
A.​ IgM antibodies are pentameric (composed of five subunits), not dimeric. This
pentameric structure gives IgM antibodies a higher avidity for antigens, but it is
not the reason for their hemolytic activity. The dimeric structure is more
associated with IgA antibodies, not IgM.
B.​ While it is true that IgM antibodies have five antigen-binding sites (due to their
pentameric structure), this fact does not directly explain their hemolytic
properties. The ability to fix complement is the crucial factor in hemolysis, not the
number of binding sites. The high number of binding sites does enhance the
antibody's ability to crosslink antigens, but it's the complement activation that
causes the hemolysis.
C.​ ABO IgM antibodies do react well at lower temperatures (4-22°C), which is why
they are known as cold-reactingantibodies. However, their hemolytic properties
are not due to their reactivity at these temperatures. Their primary hemolytic
action is through the activation of complement, not temperature-dependent
immune reactivity. Additionally, ABO IgM antibodies typically don’t react as
strongly at 37°C, the body temperature, compared to lower temperatures.
D.​ The hemolytic nature of ABO IgM antibodies is mainly due to their efficient
ability to fix complement, which leads to the lysis of red blood cells. This
makes d the correct answer. The other options (a, b, c) are incorrect because
they focus on structural characteristics or reactivity that do not directly explain the
complement-mediated hemolysis caused by ABO IgM antibodies.

36. All of the following are antigens under ISBT 001, except:
a. A
b. B
c. AB
d. H

ANSWER:
A.​ The A antigen is a well-known antigen in the ABO blood group system, and it is
an antigen under ISBT 001. So, this option is incorrect.
B.​ Similarly, the B antigen is part of the ABO blood group system and is also listed
under ISBT 001. So, this option is also incorrect.
 C.​ AB is not an antigen in itself; it refers to the presence of both A and B antigens
together on the same RBC. Thus, AB is a blood group, not an individual antigen.
This means the AB blood group does not count as a standalone antigen under
ISBT 001, so this option is also incorrect.
D.​ The H antigen is actually an important precursor for the A and B antigens in the
ABO blood group system and is classified as a separate antigen under ISBT 001.
Therefore, H is an antigen and the correct answer to the question since it is not
excluded from the list.

37 Which is correct pair for ABH Ag and its immunodominant sugar?

a. A Ag = N-acetyl-glucosamine
b. B Ag = L-fucose
c. H Ag = N-acetyl-galactosamine
d. none of the choices

ANSWER:
A.​ Incorrect. The A antigen contains N-acetylgalactosamine as the
immunodominant sugar, not N-acetyl-glucosamine. The A antigen is formed
when N-acetylgalactosamine is added to the H antigen structure.
B.​ Incorrect. The B antigen has D-galactose as its immunodominant sugar, not
L-fucose. The B antigen is formed when D-galactose is added to the H antigen
structure.
C.​ Incorrect. The H antigen has L-fucose as the immunodominant sugar. The H
antigen is a basic precursor that can be modified by the addition of either
N-acetylgalactosamine (for A antigen) or D-galactose (for B antigen).
D.​ Correct. None of the provided pairs are correct. The immunodominant sugars for
the ABH antigens are: A antigen: N-acetylgalactosamine B antigen:
D-galactose H antigen: L-fucose

38. Which of the following reagents can be used to differentiate Type O and On phenotype?
a. Al cells
b.O cells
c. Autocontrol
d. Dolichos biflorus

ANSWER:
A.​ Incorrect. Al cells are used to differentiate between A and O types, particularly for
distinguishing A from O based on the presence of A antigens, not for
distinguishing Type O from the On phenotype. Therefore, this is not the correct
reagent for differentiating between Type O and On phenotype.
 B.​ Correct. O cells are Type O red blood cells, and they contain the H antigen. The
On phenotype has weak or absent H antigen expression. By testing a sample
with O cells, one can observe a difference in agglutination or reaction based on
the strength of the H antigen expression. Type O cells (with normal H antigen)
will react differently compared to On cells, which have reduced or weak H antigen
expression. This makes O cells the correct reagent to differentiate between Type
O and the On phenotype.
C.​ Incorrect. Autocontrol refers to a test where a person's own serum is mixed
with their own red blood cells to check for autoantibodies or irregular reactions. It
does not differentiate between Type O and the On phenotype, so it is not useful
for this purpose.
D.​ Incorrect. Dolichos biflorus is a plant lectin that is used to detect the presence
of the A antigen (in A and AB phenotypes) because it agglutinates specifically
with A antigens. It is not used to differentiate Type O from On phenotype, as it
does not directly interact with the H antigen or the changes in expression
between O and On.

39. Which of the following eliminates chances of paternity? Mother: O Child: B

1. A 2. AB 3. B 4.O
a. 1 and 2
b. 2 and 3
c. 1 and 4
d. 2 and 4

ANSWER:
A.​ The father with blood type A could pass an O allele, and if the child inherited a B
allele from the mother, a B blood type could result. Therefore, 1 and 2 are not
eliminated.
B.​ Blood type AB can have a child with blood type B, so this option is incorrect
C.​ is the only correct answer because both fathers with blood types A and O are
ruled out as possible biological fathers.
D.​ As we saw, a father with blood type O cannot have a child with blood type B, but
a father with blood type AB can, so this option is also incorrect.

40. Which of the following statements is TRUE regarding the A2 blood group?
a. Dolichos biflorus lectin agglutinates A2 but not Al RBCs
b. Ulex europaeus lectin will give stronger reactions with Al than with A2 RBCs
c. A2 RBCs have more H antigen than AI RBCs
d. Most A2 individuals have a different form of anti-B than A lindividuals

ANSWER:
 A.​ There’s a substance called Dolichos biflorus lectin (DBA), which sticks to A1 red
blood cells more than to A2 red blood cells. It doesn't just stick to A2 and ignore
A1. So, this statement is false.
B.​ Another substance called Ulex europaeus lectin (UEA) actually sticks to A2 red
blood cells more than to A1 cells. This happens because A2 has more H
antigen, and UEA likes to attach to it. So, this statement is also false.
C.​ A2 red blood cells have more H antigen than A1 red blood cells. This is the
main difference between the two types of A blood. So, this statement is true.
D.​ People with both A1 and A2 blood usually have the same kind of anti-B
antibodies, which fight against the B blood type. There’s no real difference
between the anti-B antibodies of A1 and A2 people, so this statement is false.

40. Which of the following statements is TRUE regarding the A2 blood group?
a.​ Dolichos biflorus lectin agglutinates A2 but not A1 RBCs
b.​ Ulex europaeus lectin will give stronger reactions with Al than with A2 RBCs
c.​ A2 RBCs have more H antigen than Al RBCs
d.​ Most A2 individuals have a different form of anti-B than A1 individuals

a. Dolichos biflorus lectin agglutinates A2 but not A1 RBCs – Incorrect: Dolichos biflorus
agglutinates A1 cells, not A2.
b. Ulex europaeus lectin will give stronger reactions with A1 than with A2 RBCs –
Incorrect: Ulex europaeus reacts with H antigen, which is more abundant on A2 than A1
cells, so it reacts more strongly with A2.
c. A2 RBCs have more H antigen than A1 RBCs – Correct: A2 cells convert less H antigen
to A antigen, so they retain more H antigen than A1 cells.
d. Most A2 individuals have a different form of anti-B than A1 individuals – Incorrect: Most
A2 individuals do notproduce anti-B; only a small percentage produce anti-A1, not a
different form of anti-B.

41. Most common principle employed to determine ABO secretor status:

a.​ Hemagglutination - tube type
b.​ Hemagglutination - gel type
c.​ Agglutination inhibition
d.​ Autoadsorption

Hemagglutination – tube type – ❌ Incorrect: This method is commonly used for blood typing
but not typically used to detect secretor status, which involves soluble antigens.

Hemagglutination – gel type – ❌ Incorrect: Like tube testing, gel hemagglutination is used
for cell-based antigen-antibody reactions, not soluble antigen detection.
Agglutination inhibition – ✅ Correct: Agglutination inhibition is the most common
method for determining secretor status, as it detects soluble ABO antigens in saliva by
inhibiting agglutination of indicator cells.

Autoadsorption – ❌ Incorrect: Autoadsorption is used to remove autoantibodies from

patient serum, and is not related to testing for secretor status.

42. What antigens can be expressed in secretions if these genes (Hh, BO, sese) are present?
a.​ B antigen only
b.​ H antigen only
c.​ B and H antigens
d.​ none

a. B antigen only – Incorrect: The sese genotype means the person is a non-secretor and
cannot secrete any ABO-related antigens, including B.
b. H antigen only – Incorrect: Even though the H gene is present, sese prevents
secretion of H antigen.
c. B and H antigens – Incorrect: Both B and H antigens require a functional Se gene to be
present in secretions.
d. None – Correct: The sese genotype means the person is a non-secretor, so no ABO or
H antigens will be present in secretions, regardless of their ABO or Hh genotype.

43. A blood donor has the genotype ih, AB, sese, what is his apparent red cell phenotype during
routine forward and reverse group typing?
a.​ A
b.​ B
c.​ O
d.​ AB

a. A – ❌ Incorrect: Although the person carries the A allele, they cannot express A antigen
on their red cells due to the lack of H antigen.

b. B – ❌ Incorrect: Similar to A, the person has the B allele, but without H antigen, B antigen
can't be expressed on the red cells either.

c. O – ✅ Correct: Since no A, B, or H antigens are expressed on red cells in the Bombay

phenotype, forward grouping mimics type O, even though the genotype is AB.

d. AB – ❌ Incorrect: This is what the genotype suggests, but without H antigen, AB antigens
can't be formed, so the red cells won’t react as AB in testing.
44. Most probable condition causing the discrepancy in ABO typing:
Anti-A = 0
A1 cells = 0
O cells = 0
Anti-B =4+
B cells = 0
Autocontrol = 0
a.​ Cold agglutinin
b.​ Chimerism
c.​ Hypogammaglobulinemia
d.​ Leukemia or Lymphoma

a. Cold agglutinin – Incorrect: Would likely cause unexpected positive reactions at room
temp, especially with O cells or autocontrol, not missing antibodies.
b. Chimerism – Incorrect: May cause mixed-field or unexpected dual cell populations, not
an isolated absence of antibodies.
c. Hypogammaglobulinemia – Correct: This condition leads to decreased or absent
antibody production, explaining the lack of anti-A and anti-B in the reverse type.
d. Leukemia or Lymphoma – Incorrect: Can cause weak antigen or antibody expression,
but not as isolated or specificas in hypogammaglobulinemia.

45. Most probable condition causing the discrepancy in ABO typing:

Anti-A = 4+
Al cells = 1+
O cells = 0
Anti-B = 0
B cells = 4+
Autocontrol = 0

a.​ Cold agglutinin

b.​ Al subgroup with Anti-H
c.​ A2 subgroup with Anti-Al
d.​ A+ weak subgroup with Anti-B

Cold agglutinin – ❌ Incorrect: Would usually show broad, nonspecific reactivity, especially
with O cells and positive autocontrol, which are both negative here.

A1 subgroup with Anti-H – ❌ Incorrect: A1 individuals have low H antigen and don’t typically
make anti-H; also, O cells (rich in H) show no reaction, ruling this out.
A2 subgroup with Anti-A1 – ✅
Correct: A2 individuals lack A1 antigen and can produce
anti-A1, which reacts weakly with A1 cells, explaining the 1+ reverse reaction and the
discrepancy.

A+ weak subgroup with Anti-B – ❌ Incorrect: This would imply presence of anti-B, but Anti-B
= 0 and B cells react strongly (4+), indicating anti-B is clearly present, not absent.

46. Most probable condition causing the discrepancy in ABO typing:

Anti-A = 4+
Al cells = 1+ (15°C)
0 cells = 1+ (15°C)
Anti-B = 4+
B cells = 1+ (15°C)
Autocontrol = 1+ (15°C)
a.​ Cold agglutinin
b.​ Chimerism
c.​ Hypogammaglobulinemia
d.​ Leukemia or Lymphoma

Cold agglutinin – ✅Correct: Cold agglutinins react at or below room temp, cause
nonspecific weak agglutinationwith all reverse cells, and positive autocontrol, matching this
pattern.

Chimerism – ❌ Incorrect: Would typically show mixed-field agglutination, which is not

evident here.

Hypogammaglobulinemia – ❌ Incorrect: Would show absent or weak antibody responses

in reverse typing, but not explain positive autocontrol or reaction with O cells.

Leukemia or Lymphoma – ❌Incorrect: May weaken antigen/antibody expression but not

cause broad cold reactivity or autocontrol positivity like seen here.

47. If the patient is blood type O, he can only receive RBCs from blood type/s:
1.​ A
2.​ O
3.​ AB
4.​ B

a.​ 1 and 2
b.​ 2 and 3
c.​ 1,2 and 4
d.​ 2 only
1 and 2 – ❌ Incorrect: Type A is not acceptable.
2 and 3 – ❌ Incorrect: Type AB is not acceptable.

1, 2 and 4 – ❌ Incorrect: Only type O is acceptable; A and B are not.

2 only – ✅ Correct: Only type O RBCs are safe for a type O patient.

48. If the patient is blood type A, he can only receive plasma components from blood type/s:
1. A
2. O
3. AB
4. B

a.​ 1 and 2
b.​ 1 and 3
c.​ 1,2 and 4
d.​ 1 only

1 and 2 – Incorrect: O plasma is unsafe.

1 and 3 – Correct: Only A and AB plasma are safe for type A recipients.
1, 2, and 4 – Incorrect: Includes unsafe plasma types.
1 only – Incorrect: AB plasma is also safe and ideal.

49. Which of the following can resolve the ff ABO discrepancy?

Anti-A = 4+
Al cells = 2+
0 cells = 0
Anti-B = 0
B cells = 4+
Autocontrol = 0
1. Secretor studies using saliva
2. Use of acetic anhydride
3. Use of acidified reagent
4. Prewarming test components at 37C

a.​ 1 and 3
b.​ 2 and 3
c.​ 1, 2 and 3
d.​ 1 only
 ❌ Incorrect: Acetic anhydride is not useful in resolving anti-A1 discrepancies.
a. 1 and 3 –

b. 2 and 3 – ❌ Incorrect: Acetic anhydride is not helpful for resolving the ABO typing
discrepancy.

c. 1, 2 and 3 – ✅ Correct: Secretor studies and acidified reagents can help resolve ABO
discrepancies, including those involving anti-A1 antibodies.

d. 1 only – ❌ Incorrect: Acidified reagents are also helpful, so 1 only is not the best answer.
50. Cells carrying a weak-D antigen require the use of what test to demonstrate its presence?
a. DAT
b. IAT
c. Elution test
d. Microplate test

a. DAT (Direct Antiglobulin Test) – Incorrect: DAT detects antibodies already bound to red
cells in vivo, not weak D antigen expression.
b. IAT (Indirect Antiglobulin Test) – Correct: IAT is used to detect weak D antigen by
incubating patient RBCs with anti-D, then adding AHG to reveal weak or incomplete
reactions.
c. Elution test – Incorrect: Elution is used to remove and identify antibodies from
sensitized RBCs, not for detecting weak D.
d. Microplate test – Incorrect: While this is a method of testing, it is not specific to
detecting weak D and typically not the standard approach.

51. Which of the following Rh antigens is the least immunogenic?

a.​ D
b.​ C
c.​ c
d.​ e

D– ❌ Incorrect: The D antigen is the most immunogenic of the Rh antigens and is a major
factor in Rh incompatibility and hemolytic disease of the newborn (HDN).

C– ❌ Incorrect: The C antigen is highly immunogenic, though slightly less than the D antigen,
but still capable of triggering significant immune responses.

c– ❌ Incorrect: The c antigen is also immunogenic but generally less so than C and D, but
still can lead to sensitization.

e– ✅ Correct: The e antigen is the least immunogenic of the Rh antigens, meaning it is less
likely to stimulate an immune response compared to D, C, or c.
52. Weak D phenotype involving the C allele which is in trans to D allele:

A.​ Genetic Weak D

B.​ Gene interaction effect
C.​ Partial D
D.​ Rh null

RATIONALE:

A.​ This results from point mutations within the RHD gene, altering D antigen expression —
not caused by the C gene in trans.
B.​ This occurs when a person inherits a D gene on one chromosome and a C gene on the
opposite (trans) chromosome — for example: Dce/cDe.
C.​ Refers to missing or altered epitopes on the D antigen, often due to gene
rearrangements or hybrid genes. These individuals can make anti-D if exposed to
normal D antigens.
D.​ A rare condition where no Rh antigens (D, C, E, c, e) are expressed at all due to
mutations or deletions in the RH or RHAG genes. It is unrelated to Weak D due to C in
trans.

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

53. Weak D test was performed and a saline control was included in the testing. If the patient’s
red cells results to 1+ and the control tube has no reaction, how would you interpret the result?

A.​ Rh D Positive
B.​ Rh D Negative
C.​ Test should be repeated and the control tube omitted
D.​ Invalid result and Rh type cannot be determined

RATIONALE:

A.​ When performing a Weak D test, a saline control is included to ensure that any observed
agglutination is due to specific anti-D reactivity, not nonspecific factors (like
autoagglutination or protein artifacts).
B.​ Incorrect, because agglutination (1+) with anti-D indicates the presence of the D antigen.
C.​ The control tube is necessary in weak D testing.
D.​ This is incorrect because the test is legitimate with sufficient control.

REFERENCE

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.
54. Most common Rh haplotype in Asia

A.​ DcE
B.​ DCe
C.​ Dce
D.​ dCe

RATIONALE:

a.​ This haplotype is less common than DCe and dce, with a frequency of around
21%.
b.​ This haplotype is the most common in Asians, with a frequency of around 70%.
c.​ The second most common haplotype in Asians, with a frequency of around 3%.
d.​ Very rare globally, especially in Asians

REFERENCE:

Daniels G. (2013). Human Blood Groups, 3rd ed.

55. What Rh antigens are present on an individual with the phenotype R1r’

A.​ D, d, C, c, e
B.​ D, C, c, E, e
C.​ D, C, c, e
D.​ D, C, e

RATIONALE:

a.​ Individuals are either D or d (Rh-negative), not both.

b.​ E is not present in either haplotype.
c.​ D: Present (from R₁), C: Present (from both), c: Present (from r′) e: Present (from
both), E: Absent (not present in either haplotype)
d.​ Missing c, which is present from r′ haplotype.

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

56. Anti-c will not react with:

a.​ R1 R2
b.​ R2 r
 c.​ Ro r’’
d.​ Rz r’

RATIONALE:

A.​ R1 R2 = DCe / DcE

DCe: c-negative​
DcE: c-positive​
One allele has c antigen, so Anti-c will react

B.​ R2 r = DcE / dce

DcE: c-positive​
dce: c-positive​
Both have c antigen → Anti-c will react

C.​ Ro r″ = Dce / dcE

Dce: c-positive​
dcE: c-positive​
Anti-c will react

D.​ DCE: no c antigen​

dCe: no c antigen​
Both are c-negative → Anti-c will NOT react​

REFERENCE:

Harmening, D.M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.​

57. Blood factors present: Rho hr’ hr’’ / Rho rh’ hr’’

a.​ Ro R1
b.​ R1 ry
c.​ Rz r’’’
d.​ r’ R2

RATIONALE:

A.​ 1st Haplotype: Rho hr′ hr″

 D, c, e​
This is the Ro haplotype (Dce)​

2nd Haplotype: Rho rh′ hr″

D, C, e​
This is the R1 haplotype (DCe)

B.​ DCe / dCE → ry has no D, but both haplotypes are D-positive.​

C.​ DCE / dcE → Rz has E, but the given factors do not include E.​

D.​ dCe / DcE → First haplotype is Rh-negative (r′), but both in the question are
D-positive.

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

58. What percentage of their offspring will be Rh negative? Mother: R2 r ; Father rr’

A.​ 25%
B.​ 50%
C.​ 75%
D.​ 100%

RATIONALE:

A.​ If only 1 out of 4 offspring were Rh-negative, then we'd only have rr and the rest
Rh-positive. But in this case, 2 out of 4 (r′r and rr) are Rh-negative.
B.​

Offspring Genotype Rh status

R2r’ Rh-positive

R2r Rh-positive

r’r Rh-negative

rr Rh-negative

So, 2 out of 4 = 50% are Rh-negative.

 C.​ That would mean 3 out of 4 offspring are Rh-negative. But based on the Punnett
square, only 2 of 4 combinations lack the D antigen (making them Rh-negative).
D.​ If all offspring were Rh-negative, both parents would need to be homozygous
recessive for Rh (i.e., rr or r′r′). But the mother is heterozygous (R2r), so she can
pass on a D (Rh+) allele. Therefore, some offspring will be Rh-positive.

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

59. Which red cell antigen is not intrinsic to the RBC membrane?

A.​ Gerbich
B.​ Cartwright
C.​ Scianna
D.​ Chido-Rogers

RATIONALE:

A.​ Intrinsic to RBC membrane. Located on glycophorin C and D (part of RBC

structure)​

B.​ Also known as Yt antigen. Found on acetylcholinesterase, which is embedded in

the RBC membrane
C.​ Antigens are carried on ERMAP protein, a transmembrane protein
D.​ Not intrinsic to the RBC membrane. Antigens are plasma-derived complement
fragments (C4) that adsorb onto the RBC surface. They are adsorbed, not
produced by the RBC itself

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

60. Red cell antigens of this blood group system are absent in cases of PNH:

A.​ Chido-Rogers
B.​ Gerbich
C.​ Cromer
D.​ Knops

RATIONALE:
 A.​ These antigens are not intrinsic—they’re adsorbed complement fragments (C4)
from plasma. Not affected in PNH.
B.​ Antigens found on glycophorin C and D, which are integral membrane proteins,
not GPI-anchored. Remain present in PNH.
C.​ Cromer antigens are carried on DAF (Decay-Accelerating Factor / CD55), which
requires a GPI anchor. CD55 is absent or deficient in PNH, so Cromer antigens
are also absent.
D.​ Located on CR1 (Complement Receptor 1)—an integral protein, not
GPI-anchored. Present in PNH.​

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

61. Anti-PP1Pk is produced by individuals with this phenotype early in life without RBC
sensitization:

A.​ P1
B.​ P2
C.​ P1k
D.​ p

RATIONALE:

A.​ Has the P1 and P antigens. Will not produce anti-PP1Pk.

B.​ Lacks P1, but still has P antigen. May make anti-P1, not anti-PP1Pk.
C.​ Has P1 and Pk, lacks P. May make anti-P, not anti-PP1Pk.
D.​ Has none of the P1, P, or Pk antigens (i.e., the p phenotype). Can produce
anti-PP1Pk naturally.

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

62. Autoantibody in Paroxysmal Cold Hemoglobinuria described as a biphasic

hemolysin:

A.​ IgG
B.​ IgM
C.​ IgA
D.​ IgD
RATIONALE:

A.​ In Paroxysmal Cold Hemoglobinuria (PCH), the autoantibody involved is a

biphasic hemolysin known as the Donath-Landsteiner antibody, which is IgG in
nature.
B.​ Although IgM is commonly cold-reactive (as in Cold Agglutinin Disease), PCH is
specifically mediated by IgG, not IgM.
C.​ Rarely implicated in hemolytic processes; not involved in PCH
D.​ Not typically involved in hemolytic anemia or autoantibody formation.

REFERENCE:

Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices, 7th ed.

75. Positive result in AHG phase of antibody screen and a negative result in autologous control
suggests:
A.​ Presence of autoantibody, IgM type - Autoantibodies generally react in the autologous
control. Since this result is negative, the cause is likely not an autoantibody.
B.​ Presence of alloantibody, IgM type - IgM alloantibodies would react at room temperature,
usually in the immediate spin phase, not the AHG phase.
C.​ Presence of alloantibody, IgG type - IgG alloantibodies cause reactions in the AHG
phase and can result in a negative autologous control, indicating the presence of an
alloantibody.
D.​ Presence of autoantibody, IgG type - IgG autoantibodies would typically show reactivity
in both the AHG phase and autologous control, making this less likely.
Reference: Harmening, D. M. (2019). Modern blood banking and transfusion practices (7th ed.).
F.A. Davis Company.

76. If a recipient is incompatible with only one donor, one should suspect:
A.​ ABO discrepancy - An ABO discrepancy would usually cause incompatibility with
multiple donors, not just one.
B.​ clerical error - typically affect multiple donors, not just one. The scenario points to
something more specific, like an antigen reaction.
C.​ recipient antibody reacting to a low incidence antigen - Low incidence antigens are rare,
so the recipient’s antibody could react with a single donor who expresses that rare
antigen, causing incompatibility.
D.​ recipient antibody reacting to a high incidence antigen - High incidence antigens are
present on most red blood cells, so antibodies to these would cause incompatibility with
multiple donors, not just one.
Reference: Harmening, D. M. (2019). Modern blood banking and transfusion practices (7th ed.).
F.A. Davis Company.
77. Next step when a clinically significant antibody is identified from the patient sample:
A.​ antigen-positive blood units should be chosen for crossmatch - Choosing
antigen-positive units would result in incompatibility with a clinically significant antibody,
leading to potential transfusion reactions.
B.​ antigen-negative blood units should be chosen for crossmatch - After identifying a
clinically significant antibody, the next step is to select antigen-negative blood that lacks
the specific antigen to prevent positive crossmatch and ensure transfusion safety.
C.​ perform elution to dissociate antibodies - Elution is not the next step for a clinically
significant antibody after identification. It is used for cases like positive DAT or when
investigating antibodies already bound to red cells.
D.​ perform an electronic crossmatch - Electronic crossmatch is typically used when no
clinically significant antibodies are present. For a patient with clinically significant
antibodies, selecting antigen-negative units is critical.
Reference: Harmening, D. M. (2019). Modern blood banking and transfusion practices (7th ed.).
F.A. Davis Company.

78. Antibody screening done on a patient shows positive reaction pattern on all three screening
cells in the AHG phase. What is the most likely reason for this result?
A.​ An alloantibody against a high-frequency antigen is present - High-frequency antigens
are present on most red blood cells, so an alloantibody targeting one could cause
reactions with all screening cells.
B.​ Mixture of different alloantibodies are present - A mixture of alloantibodies could also
explain the positive result on all three screening cells, as each cell could have different
antigens that the antibodies can react with.
C.​ Either of the choices - Correct. Either the presence of a high-frequency antigen antibody
or a mixture of different alloantibodies can cause positive reactions with all three
screening cells.
D.​ None of the choices - The positive reaction pattern can indeed be explained by either a
high-frequency antigen antibody or a mixture of alloantibodies, so this option is not
applicable.
Reference: Harmening, D. M. (2019). Modern blood banking and transfusion practices (7th ed.).
F.A. Davis Company.

79. What should be the next step if a patient currently has a negative antibody screening but
turn out to have a blood bank history of possessing anti-Jk^a?
A.​ Release type O Rh D negative red cells only - The patient’s anti-Jk^a antibody can react
with red cells positive for Jk^a, regardless of Rh D status, making this option insufficient.
B.​ Crossmatch using random blood units, since antibody screening is negative - A negative
antibody screen doesn’t rule out the presence of an anti-Jk^a antibody, which requires
testing with Jk^a-negative blood.
C.​ Perform an electronic or computer crossmatch if patient has a record of at least two
blood typing results - An electronic crossmatch alone doesn’t account for antibodies like
anti-Jk^a. Physical crossmatch with Jk^a-negative units is necessary.
 D.​ Perform an IS and AHG crossmatch using blood units negative for Jk^a antigen -
Correct. Since the patient has anti-Jk^a, the crossmatch should be performed with
Jk^a-negative blood to ensure compatibility. Both Immediate Spin (IS) and AHG
crossmatches are required to check for IgM and IgG antibodies.
Reference: Harmening, D. M. (2019). Modern blood banking and transfusion practices (7th ed.).
F.A. Davis Company.

80. Which is correct about the Antibody Panel cells?

A.​ At least 2 or 3 vials are needed when performing antibody identification - An antibody
panel should have 2 or 3 vials of group O red cells with varying antigen profiles for
accurate antibody identification.
B.​ It should have the same pattern of antigen expression for all vials - as the panel needs a
range of antigen profiles to distinguish between different antibodies.
C.​ It should include double dose of Rh, Kidd, MNS and Duffy antigens - as double doses of
antigens are not necessary; the focus is on having diverse antigens.
D.​ Made up of Type O red cells phenotyped with loss commonly encountered antigens - as
the cells should contain a broad range of antigens, not the loss of common ones.
Reference: Harmening, D. M. (2019). Modern blood banking and transfusion practices (7th ed.).
F.A. Davis Company.

81. You have performed an antibody screen using the tube method. All three screening cells
tested negative. The Coombs check cells in all three tubes are also nonreactive. What should
you do?

A.​ Respin the tubes and reread them - may not resolve the underlying problem, performing
an antibody identification panel
B.​ Perform an antibody identification panel - is unnecessary without a positive screen, and
crossmatching
C.​ Repeat the antibody screen - when Coombs check cells are nonreactive, as this
suggests an issue with the AHG phase of the test.
D.​ Perform the next step: crossmatch - should only be done after confirming a valid
antibody screen result.
Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices (7th
ed.).​

82. Identify the reacting antibody in the antibody panel

 A.​ Anti-M - Reacts with cells that have the M antigen in the MNS blood group system,
typically at room temperature, and is usually an IgM antibody.
B.​ Anti-c - Reacts with the c antigen in the Rh system, usually at 37°C or AHG phase, and
is an IgG antibody.
C.​ Anti-Fy" - the Duffy blood group system, typically seen in Fy(a-b-) individuals, and is
usually an IgG antibody, clinically significant for transfusions and HDN.
D.​ Anti-D - Reacts with the D antigen in the Rh system, usually an IgG antibody, and is
significant in Rh incompatibility, HDN, and hemolytic transfusion reactions.
Reference: Harmening, D. M. (2019). Modern Blood Banking and Transfusion Practices (7th
ed.).

83. Minimum required hemoglobin level for allogeneic blood donation:

A.​ 125 g/L - common recommendation in blood donation for healthy donors, but the
minimum can vary between 110 g/L (as required by many guidelines) and 125 g/L
(recommended for some populations, particularly females).
B.​ 110 g/L - This is typically the minimum hemoglobin level for safe donation in most
guidelines.
C.​ 100 g/L - Below the safe minimum for donation; usually not allowed.
D.​ 115 g/L - This is a valid hemoglobin level but is not the universally defined minimum.
Reference: American Red Cross. Donor Eligibility Guidelines.​

84. Potential Donors who have had a history of malaria should be:
A.​ permanently rejected - This is not the standard recommendation unless the person has
ongoing chronic malaria infection or recurrent episodes, which is uncommon. Most
people recover fully from malaria with treatment.
B.​ deferred for 6 months - This is a shorter deferral period than what is recommended for
malaria. Some guidelines specify 12 months, not 6 months.
 C.​ deferred for 1 year after cessation of treatment - This is the most commonly
recommended deferral period. After completing treatment for malaria, most blood
donation guidelines recommend waiting for a period of 1 year before donating to ensure
that any potential residual malaria parasites have cleared.
D.​ deferred for 3 years after cessation of treatment - helps minimize the risk of transmitting
malaria parasites, especially considering the possibility of relapsing malaria, which could
be asymptomatic but still infectious.
Reference: World Health Organization (WHO). Blood Donor Selection: Guidelines on Assessing
Donor Suitability.

85. Optimal diastolic blood pressure for blood donation:

A.​ 90-160 mmHg - This range is too high for diastolic blood pressure and may indicate
hypertension, which could disqualify someone from donating blood.
B.​ 90-120 mmHg - While within normal limits, a diastolic pressure lower than 90 mmHg may
not meet the higher threshold required for blood donation.
C.​ 60-90 mmHg - This is the optimal range for diastolic blood pressure when donating
blood, ensuring the donor is in good cardiovascular health.
D.​ 40-50 mmHg - This range is too low and would indicate hypotension, which could
disqualify a donor as it could pose a risk for fainting or other complications.
Reference: American Red Cross (2022). Blood Donation Eligibility Requirements.

86. Platelet concentrate units should have a pH of:

A.​ ≥6.2 - The pH requirement for platelet concentrates to ensure they are functional and
viable during storage and transfusion.
B.​ ≤ 6.2 - This would indicate an improper pH level for platelet storage, which could lead to
a decrease in platelet function and quality.
C.​ ≥ 7.4 - This pH is within the normal blood pH range but is too high for platelet storage; a
pH above 7.4 could harm the platelets.
D.​ ≤7.4 - This would imply a pH closer to neutral but still outside the optimal range for
platelet preservation.
Reference: American Association of Blood Banks (AABB). (2016). Standards for Blood Banks
and Transfusion Services. 29th edition.

87. A single whole blood derived platelet concentrate contains a minimum of platelets:

a: 5.5 x 10'0. c. 3.0 x 1010

b. 5.5 x 1011. d. 3.0 x 1011

Answer:

a.​ Minimum standard for platelets from one unit of whole blood.
b.​ Too high — typical for apheresis platelets, not whole blood-derived.
c.​ Below apheresis level; above whole blood standard.
 d.​ Higher than expected for whole blood-derived; fits apheresis units.

reference:

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

88. FFP should be thawed at a temperature of for preparation of cryoprecipitate:

a. 30°C. c. 4°C

b. 37°C. d. -18C

Answerk

a.​ Too warm; not used for cryoprecipitate preparation.

b.​ Used to thaw FFP for transfusion, but not when preparing cryo.
c.​ FFP is thawed at 1–6°C to allow precipitation of cold-insoluble proteins (Factor VIII,
fibrinogen, vWF), which are collected as cryoprecipitate.
d.​ Storage temperature for frozen FFP, not thawing.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

89. Leukoreduced RBC units must have a residual content of leukocytes.

a.​ <5 × 10^9. c. <3 × 10^6

b.​ <5 × 10^6. d. <3 × 10^11

answer

a.​ Too high — typical of non-filtered units.

b.​ Acceptable upper limit of leukocytes in leukoreduced RBCs as per minimum standard.
c.​ Stricter than required — this is a target for prestorage leukoreduction.
d.​ Far above standard — not acceptable for leukoreduced units.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

90. As part of QC, Granulocyte concentrate units should contain granulocytes

a. <5 × 10^6 c. <1 × 10^10

b. >5 × 10^6 d. >1 × 10^10

answer
 a.​ Too low — not acceptable for therapeutic use.
b.​ Still below standard therapeutic dose.
c.​ Insufficient — doesn't meet QC requirements.
d.​ Minimum granulocyte content required per AABB standards for granulocyte
concentrates.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

91. FFP is plasma collected from whole blood units which is frozen within:

a. 4 hours. c. 12 hours

b. 8 hours. d. 24 hours

answer

a.​ Too early — not the standard cutoff.

b.​ FFP must be frozen within 8 hours of whole blood collection to preserve clotting factors.
c.​ Too late — used for plasma frozen within 24 hours, not FFP.
d.​ Used for PF24, not standard FFP.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

92. A patient has a hemoglobin of 8 g/dL. The surgeon wants to raise the hemoglobin to 10 g/dL
before surgery. How many units of RBC need to be administered to this patient to raise the
hemoglobin to the required level?

a.​ 1
b.​ 2
c.​ 3
d.​ 4

answer

a.​ Not enough; 1 unit generally raises hemoglobin by 1 g/dL, so more is needed.
b.​ 2 units of RBCs should raise the hemoglobin from 8 g/dL to 10 g/dL.
c.​ Too many; 2 units should be sufficient for the desired hemoglobin increase.
d.​ Excessive; 2 units are enough to achieve the target hemoglobin increase.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

93. A patient has a platelet count of 15,000/uL, and there are no complicating factors such as
fever or HILA sensitization. If he is given a platelet pool of 6 units, what would you expect his
post-transfusion count to be?

a. 21,000-27,000/uL. c. 45,000-75,000/uL

b. 25,000-35,000/uL. d. 75,000-125,000/L

answer

a.​ Too low, not a reasonable range for 6 units of platelets.

b.​ This is a bit low; 6 units of platelets should raise the count more.
c.​ Typically, 6 units of platelets will raise the count to this level in a patient with a starting
count of 15,000/uL.
d.​ Too high; this would be more realistic with more units of platelets or if the patient's
response is unusually high.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

94. According to AABB, Cryoprecipitate should contain at least of fibrinogen and. of Factor VIII.

a. 250 mg; 160 IU c. 100 mg; 50 IU

b. 150 mg; 80 IU. d. 60 mg; 30 IU

answer

a.​ Too high — exceeds the minimum standards.

b.​ Higher than the required minimum values.
c.​ Higher than the required minimum values.
d.​ This is the minimum requirement according to AABB for fibrinogen and Factor VIII
content in cryoprecipitate.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

95. In preparing platelets from a unit of whole blood, the correct order of centrifugation is:

a. Hard spin followed by a hard spin. c. Light spin followed by a hard spin

b. Light spin followed by a light spin. d. Hard spin followed by a light spin

answer
 a.​ Not correct; both spins should be of different intensities.
b.​ Not correct; platelets need to be separated from the red blood cells in a two-step
process.
c.​ The proper method for separating platelets from whole blood. The light spin first
separates the plasma, and the hard spin follows to concentrate the platelets.
d.​ Incorrect order; the hard spin should come after the light spin for optimal platelet
separation.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

96. Most cases of Febrile non-hemolytic transfusion reaction are due to Patient's triggering
release of proinflammatory cytokines

a. anti-HLA. c. anti-IgA

b. plasma protein antibodies. d. anti-leukocytes

answer

a.​ HLA antibodies can cause reactions but are less common in febrile non-hemolytic
transfusion reactions.
b.​ This is more related to allergic reactions than febrile non-hemolytic reactions.
c.​ These can cause severe reactions in patients with IgA deficiency but are not the main
cause of febrile non-hemolytic reactions.
d.​ The most common cause; antibodies against donor leukocytes trigger the release of
cytokines that cause fever.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

97. Antibodies associated with Transfusion-Related Acute Lung Injury:

a. anti-HLA. c. anti-igA

b. plasma protein antibodies. d. anti-leukocytes

answer

a.​ Anti-HLA antibodies are the primary cause of TRALI, as they can trigger an immune
response leading to lung injury.
b.​ Not the primary cause of TRALI; these antibodies are more related to allergic reactions.
c.​ These antibodies are associated with allergic reactions or anaphylaxis, not TRALI.
d.​ Though anti-leukocyte antibodies are involved in TRALI, anti-HLA antibodies are more
directly implicated in the pathogenesis of the condition.
reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

98. Measurement of BNP levels, may be helpful in the diagnosis of:

a. Anaphylactic transfusion reaction. c. Transfusion-Related Acute Lung Injury

b. Febrile non-hemolytic transfusion reaction. d. Transfusion-Associated Circulatory Overload

answer

a.​ BNP is not typically used to diagnose anaphylaxis, which is more related to
IgE-mediated hypersensitivity.
b.​ BNP is not helpful in diagnosing febrile non-hemolytic transfusion reactions, which are
primarily related to cytokine release.
c.​ BNP levels may be elevated in TRALI, but they are not the primary diagnostic marker.
d.​ BNP levels are helpful in diagnosing transfusion-associated circulatory overload (TACO),
as BNP is a marker of heart strain and volume overload.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

99. Clinically significant ABO HDFN occurs most frequently in group mothers who have a group
infant.

a. A or B; AB. c. 0; A or B

b. A or B; 0. d. 0; 0

answer

a.​ Less likely to cause ABO HDFN compared to other combinations.

b.​ While this is possible, it is less common than the correct answer.
c.​ Most common scenario for ABO HDFN, as mothers with blood type O can produce
antibodies against A or B antigens present in the infant.
d.​ This combination is less likely to cause clinically significant ABO HDFN because there is
no antigen difference between mother and infant.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).

100. Compute for # doses required of RhiG for the maternal patient:

Rosette test: (+)

Kleihauer-Betke test: 24 pink cells and 1,976 ghost cells

a. 2. c. 4

b.3. d. 5

answer

a.​ Too low for the fetal blood volume, clinical guidelines typically require more.
b.​ Correct answer. Standard practice for the estimated fetal blood volume
c.​ Too high for this case, unnecessary based on fetal blood volume.
d.​ Excessive, not recommended unless in exceptional cases.

reference

●​ Daniels, G. (2013). Human Blood Groups (3rd ed.).