DEPARTMENT OF MICROBIOLOGY AND IMMUNOLOGY

SEMESTER 3 MODULE TWO CONTINUOUS ASSESSMENT

PRACTICAL EXAMINATION (MI 201.02)
31ST MARCH 2021
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GENERAL INSTRUCTIONS
1. This examination consists of 25 spots
2. Two minutes are allocated for each spot
3. Your required to attempt all questions
4. The slide will automatically change to the next
question after every two minutes
5. Write your registration number and answers on the
provided answer sheet
 1

Mention 2 Advantages and 2 Disadvantages

of PCR
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 Advantages and disadvantages of PCR

advantages disadvantages
• 1:easy to set up • 1:extremely liable to
• 2:fast turnround time contamination
• 2:high degree of operator
skills required
 2

Mention four uses of eggs in Microbiology

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 Four uses of egg in microbiology
• 1:it is used for viral culture eg embryonated egg
• It is used to provide nutrient in some medium eg LJ
medium
3

Mention 4 laboratory techniques used in

identification of viruses
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 Laboratory techniques used in
identification of viruses
• EM….
• Serology(antigen or antibody detection)
• Virus culture-detect cytopathic effect or
antigen
• Molecular methods
-PCR
-SEQUENCING
 4

Above is the instrument used during phosphatase

What is the name of the instrument?
a) What is the principle used for that instrument?
b) How do you interpret the results
 Lovibond comparator
• Principle
Free chlorine reacts with DPD to form a red
violet compound,
by comparing the color against a set of
reference standards, the amount of free residue
chlorine can be determined
The amount of free residue chlorine in the water
should normally be greater than 0.2mg\l and
less than 0.5mg\l
 5

Provide interpretation for MPN table

provided below
Number of tubes giving MPN Interpretation,
positive reaction out of counts/100ml of coliforms not
5, 10 mls each water [Link]
satisfactory
a)1 2.2
suspicious
a)2 5.1
suspicious
a)3 9.2
unsatifactory
a)4 16.0
 6

Above is one of the methods to test for milk

sterility.
a) What is the name of the test?
b) Interpret the results on both tubes and give
conclusion
• A)Methylene blue reduction test-test in milk
• Tube 1(left);no reduction in methylene blue in
30min hance satisfactory
• Tube2(right):there is reduction of methylene
blue within 5minutes hence the milk is
unsatisfactory
 7

Above is one of the tests used in public health

bacteriology
a) What is the name of the test?
b) How do you determine the degree of
contamination in this method?
c) Mention one other test that can be used to
check the suitability of milk
 Miles and misra technique
• Degree of bacteria contamination of milk may
be determined by doing total viable bacteria
counts and determining presence or absence
of fecal contamination(E-COLI)
• The methylene blue test
 8

You are provided with a photo of item above

a) Name the item
b) Name the solid phase of the item
c) Mention two immunoassay that can
be performed using the item
• A) a microtiter plate
• B)-well of microtiter
plate,beads:plastic,microbeads,nitrocellulose
paper
• C)1:ELISA
2:AGGLUTINATION
 9

1/20 1/40 1/80 1/160 1/320 1/640 Control

You are provided with tubes that were used for

performance of an immunoassay
a) Name the immunoassay
b) Mention the titer of the immunoassay
c) Mention one advantage of the immunoassay
d) Mention one disadvantage of the immunoassay
• A):Tube agglutination
• B) Titer=160
• C)adv:easy to perfom
D)disad:sometimes they have low specificity
 1/ 1024
1/ 128
1/ 256
1/ 512
1/ 16
1/ 32
1/ 64
1/ 2
1/ 4
1/ 8

Neg
Pos
10
1
2
3
4
Patient # 5
6
7
8

You are provided with immunoassay test results

a) Give the Titer of a patient number 3
b) Give the Titer of a patient number 6
c) What does the test measure?
d) Mention one disadvantage of the test
 Agglutination\hemagglutination
• A)titer=512
• B)Titer=128
• Test used to QUANTITATIVE detect antibody in
patient`s sample
• Disadv:sometimes they have low specificity
11

List parameters in CSF that are useful in

differentiating bacterial meningitis from
non-bacterial meningitis
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• 1:cell count
• 2:cell types(differential cell count)
• 3:protein content
• 4:glucose level
• 5:microscopic search for microorganisms
12

List bacteria that are mainly isolated in

neonates with bacterial meningitis
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• Group b strep([Link])
• E-coli
• Listeria monocytogenes
• [Link]
 13

You are provided with gram stain of an organism

isolated from a patient with meningitis
a) Report the findings
b) Mention two culture media useful in isolating
the organism
c) Name the pathogen
• Gram-negative coffee-bean shaped diplococcic
• 1:chocolate agar base containing VCNT
• 2:THAYER MARTIN MEDIUM
• NEISSERIA MENINGITIDIS
14

Mention 4 indicators for bacteria growth

on blood culture bottles
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• PELLICLE
• SEDIMENTS
• TURBIDITY
• FLOCULENT
 15

A stained smear from a CSF specimen collected

from a patient presenting with severe headache,
fever and convulsion
a) What is the mostly likely identity
b) Name one disease caused by organism seen
 CRYPTOCOCCUS NEOFORMAS
• DS:CRYPTOCOCCUS MENINGITIS
16

a)Mention 2 common enrichment broth

useful for isolation of enteric bacteria
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a)Mention 2 lactose and 2 non-lactose gram

negative Enterobacteriaceae
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• A)-Tetrathionate broth
-selenite broth
B)@NON-LACTOSE FERMENTER
-salmonella spp
-shigella except shigella sonnei
@LACTOSE FERMNTER
-E-coli
-klebsiella
17

a) Define selective Media

b) Mention 3 content of MacConkey agar
and function of each content
• Selective media are solid media which are prepared in such a
way that the growth of some bacteria is inhibited while
selecting for the growth of other
• 1:lactose-source of carbohydrate
• 2:bile salts and crystal violet-selective agents that inhibit
growth of gram pos organism and proliferate growth of gram
neg
• 3:neutral red- ph indicator(indicator)
18

Mention four biochemical tests that are

used to identify enteric bacteria in
Microbiology lab
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• -OXIDASE TEST
• TSI
• IMViC
• -UREASE TEST
• -INDOLE TEST
 19

a) Describe the principle of the above technique

b) What is the use of this technique in microbiology
laboratory?
• Uses:This approach is used for qualitative
detection and identification of antigens
• Principle:An extract of the organism is layered
in tubes over known antisera to the different
types. A ring of precipitation occurs at the
interface with the specific antiserum
 20

a) What causes a prozone effect?

b) What will you do to remove a prozone effect?
 Prozone effect
• Cause:antibody excess,blocking antibody,non-
specific inhibitors in serum(No visible
reaction/lattice formation)
• NB:normally ppt occur at high dilution
-at low or undiluted no visible ppt
 21

a) Describe the principle of single immunodiffusion

b) What is the use of this technique in microbiology
laboratory?
• Principle:antigen placed in a well diffuses into
agar containing a specific [Link] Ab-Ag
interaction is defined by a well defined ring of
precipitation around the Ag well
• Uses:quantative
 22

a) Why is electric current applied in

immunoelectrophoresis?
b) Mention two sources of error in
immunoelectrophoresis
Electrophoresis Sources of Error
• Applying current in wrong direction.
• Incorrect buffer pH
• Incorrect timing
• Incorrect current applied.
• Concentration of reactants must be appropriate.
Why current:Immunodiffusion can be combined with
electrical current to speed things up.
 23

You are provided with disk diffusion test for antibiotic

susceptibility testing
a) Name the method used for this test
b) Give the name of the medium used for this test
c) Mention another disk diffusion method applying
the same principle
d) How is the antibiotic susceptibility determined in
this test?
• A)disk diffusion method(Kirby bauer )
• B)medium:muller hinton
• C:giant diagnostic sensitivity method
• D) test inhibition zone is measured
-sensitive:when radius equal,wider,or not more than 3mm
smaller than control
-moderate sensitive:zone radius equal,wider or not more than
3mm, but smaller than control by more than 3mm intermediate
-resistant:radius 3mm or less
 24

You are provided with a set of test for antimicrobial

sensitivity testing
a) Name the test
b) What does the test determine?
c) Read and interpret the finding
d) Mention another test used for the same purpose
• A) tube broth dilution method for
antimicrobial sensentivity test
• B)-minimum inhibitory concentration(M.I.C)
-Minimum bactericidal
concentration(M.B.C)
C)-M.I.C=8ug/ml
---M.B.C=16ug/ml
D)other test:E-test
25

Mention four factors determining

zone sizes in agar diffusion tests
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 Factors determine zone sizes of
agar diffusion test
• medium
• Amount of inoculum
• Disk contents
• Length of incubation
• ELISA
• WESTERN BLOT
• CPE
• DECOY PARTICLE