IMMUNOHEMATOLOGY - FINALS
COMPATIBILITY TESTING
NAME OF LECTURER: Rhey Gregory Aquino - UNIVERSITY OF BAGUIO
BY: Carrie Lhee B. Boado
obsolete.
Performed to
2 IMPORTANT ASPECTS
detect any
1. PHENOTYPING unexpected
➢ Performed to determine the blood type of patients and antibodies that
donors in order to confirm the blood type prior to may have
transfusion. formed
➢ There are some cases where patients are not because of
blood-typed at all because of determining their blood alloimmunizati
types prior to them being within the confines of the on.
hospital (E.g. patients could use identification cards
that provide info about their blood type).
➢ Important in order to minimize the numbers of units that 3 PHASES OF SEROLOGICAL CROSSMATCH
you need to choose from.
➢ Most commonly used: 1. Immediate Spin
- Tube or column method: Much sensitive ● Goal: To detect any cold reacting antibodies (IgM).
than slide method ● If there is agglutination, stop. If there are no agglutinations,
➢ Comprises ABO (forward and reverse) and Rh typing. proceed to the next phase.
● MAJOR CROSSMATCH
2. CROSSMATCH ➢ Mix plasma or serum of the patient (serum is
preferable to not miss out any hemolytic reactions) +
MAJOR MINOR RCS of the donor.
➢ Use tube or gel column method.
● Detects any antibodies ● If there are any ➢ 2:1 — 2 parts of the patient's serum + 1 part of the
within our patients that antibodies within the donor's RCS.
could interact with blood donors that could react
cells of the donor to with the RBC of your ● MINOR CROSSMATCH
remove any instances of patients (REVERSE OF ➢ Most samples from the donor would be anticoagulated
reducing the survivability MAJOR) whole blood samples, thus, serum would be impossible
of blood cells. ● Considered as minor to extract so plasma is used instead.
● Most important part of because: ➢ Mix patient’s RCS + donor’s plasma.
the cross matching 1. Dilution of ➢ 2:1 — 2 parts of the patient's RCS and 1 part of the
procedure. antibodies of donor’s plasma.
the donor
once PROCEDURE:
incorporated in
the circulation 1. In a tube, add 2 drops of the patient's plasma and add 1 drop of
of patients. In the donor's RCS.
cases of 2. Shake gently. Centrifuge. Shake gently, dislodging the cell
anemia, the button. Check for agglutinations or hemolysis.
unit 3. Evaluate (grade) the reactions. Write the grade on the
transferred will worksheet. Provide interpretations when tubes yielded a
be PRBC positive reaction (agglutination/hemolysis). If no agglutinations
(minimized were seen, proceed to B.
amount of
plasma =
lower antibody
titers within 2. Thermo/37 C/Incubation Phase
donor = there ● Achieved through the use of dry cell incubator or water bath
are less ● Goal: To detect the warm-reacting antibodies (IgG).
antibody that ● For the column method, use the pre-built incubator of the gel card
would react which is attached to the centrifuge of the gel cards.
and destroy ● Enhancement medium: The use of potentiators can reduce the
the px RBC) incubation period from 30-120 mins to 30-60 mins or 10-15 mins.
2. AB screen ➢ ALBUMIN
and - Macromolecules that push together antigens
identification: closer to the antibodies thereby promoting
Makes minor agglutinations. If used, time of incubation is
crossmatch reduced to as short as 30-60 mins
➢ LISS and Polybrene
- Reduces zeta potential. Incubation time:
10-15mins
➢ PEG
- Eliminates the need to perform the incubation
phase.
- Interact and absorb water molecules around
the test medium, thus, concentrating the
serum thereby promoting antigen-antibody
interaction.
PROCEDURE:
1. For tubes that yielded a negative result in the IS phase, add
one drop of LISS in the tube. Gently shake the tube. Cover
tubes with Parafilm tape.
2. Incubate the tubes in a water bath set at 37°C. Incubate for 15
minutes.
3. Centrifuge (20 secs @ 3.400 rpm) the contents of the tube.
Gently shake to dislodge the cell button. Check for
agglutinations or hemolysis.
4. Grade the reactions. Write the grade on the worksheet. Provide
interpretations when tubes yielded a positive reaction
(agglutination/hemolysis). If no agglutinations were seen,
proceed to C.
3. AHG Phase
● Follows the principle of IAT.
● Prior to adding the AHG reagent, it is vital to wash the contents of
tubes at least 3x using NSS. This prevents neutralization of the
AHG antibodies/reagent in order to only detect antibodies that had
sensitized the RBC’s — avoid false negatives and positives.
● Goal: Detect IgG that had sensitized RBC but had not agglutinated
the RBC’s.
LAST STEP RESULTS AND
INTERPRETATION
Either add a drop of CHECK Major (-) and Minor (-)
CELLS or perform ➢ The donor (entire blood
MICROSCOPIC EXAMINATION.\ unit) is compatible with
the patient (ABO
CHECK CELLS:
identical).
➢ TYPE O SENSITIZED
WITH IgG ANTI D
Major (+) and Minor (-)
➢ Checks if the AHG is
➢ Incompatible
working properly.
➢ After addition, centrifuge Major (+) and Minor (+)
for 20 secs at 3,400 rpm ➢ Completely
then check for incompatible.
agglutinations.
Major (-) and Minor (+)
➢ NOTE: If there were no
➢ Only the RBC portion
agglutinations in the can be transfused —
AHG phase, there with caution.
should be agglutinations
after the addition of COMPATIBLE: - (NO
check cells. If there are AGGLUTINATION)
still no agglutinations,
the test is considered INCOMPATIBLE: +
invalid. (AGGLUTINATION)
MICROSCOPIC EXAMINATION
➢ Place the contents of the
tube on the glass slide
then spread as wide —
examine.