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Clinical Pathology II > Serology > Flashcards

Serology Flashcards

1
Q

serology

A

detection and measurement of antibodies or antigens

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2
Q

what is serology used for?

A

can confirm viral, bacterial, or fungal infections

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3
Q

what type of antibodies are used in serology tests?

A

monoclonal antibodies to many different antigens which help to rapidly detect disease

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4
Q

what does protein electrophoresis do? (EPH)

A

assesses cause of severe hyperproteinemia (usually from a rise in immunoglobulins)
measurement results in a graph
gamma = immunoglobulins

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5
Q

IEP

A

patient’s serum is placed in a well and antibody to whole serum in a trough
comparisons are done to normal serum which can determine if there are deficiencies in 1 or more serum components or if there is an overabundance of some serum components (based on thickness of line on graph)

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6
Q

PCR tests

A

polymerase chain reaction
DNA or RNA is amplified into millions of copies and can then be detected for analysis of hereditary diseases, forensic, paternity, or diagnosis of infectious disease

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7
Q

advantages of PCR tests

A

much more sensitive and specific, quick turn-around time

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8
Q

disadvantages of PCR tests

A

high cost, false positives from contamination, technical expertise needed

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9
Q

how are PCR tests done?

A
  1. 2 strands of DNA double helix are melted, separated, and then cooled
  2. DNA enzymes are added using DNA primers that are specific for the target DNA so they complement and are copied exponentially
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10
Q

electrophoresis

A

technique used to separate molecules of DNA, RNA, and protein based on size
larger fragments move slow, smaller fragments move fast
heavier fragments stop sooner, they move from negative to positive change

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11
Q

interpretation of electrophoresis

A

look for bands at specific markers
no bands = negative
indeterminate = bands don’t match criteria for specific disease

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12
Q

Coombs test

A

detects auto-antibodies
direct test for hemolytic disease (especially IMHA)
RBCs from patient are incubated with species-specific reagent: agglutination seen if positive
too expensive to stock in-house, usually send out

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13
Q

Rh factor

A

protein on RBC
can produce antibodies against fetus in human
associated with Coombs test

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14
Q

sensitivity

A

true positives
proportion of animals with a positive result actually having the disease

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15
Q

specificity

A

true negatives
proportion of animals with a negative test that don’t have the disease

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16
Q

ELISA tests

A

enzyme-linked immunosorbent assay
types: direct, indirect, sandwich, and competition/blocking
involves lots of pipetting and washing
some are available as kits

17
Q

direct ELISA

A

quick
can be opposite: looking for antigen in sample instead of antibodies

18
Q

how do you perform a direct ELISA?

A
  1. antigen is affixed to the well surface
  2. patient sample is added (already linked with enzyme)
  3. if antibody is in the sample it will bind and form a complex
  4. wash
  5. read color change
19
Q

ELISA SNAP tests vs rapid tests

A

similar specificity and sensitivity
ELISA includes washing step
rapid tests have more potential for false positives due to not having a washing step
ELISA tests are gold standard
send out for confirmation when there is a positive rapid test

20
Q

lateral flow tests

A

a type of rapid test
immunochromatography assay (ICT)
uses colloidal gold instead of enzymes
gold buildup as sample migrates which can lead to a positive result

21
Q

indirect ELISA

A

antigen is added to wells and then patient sample is added (if the antibodies for the antigen are present it will bind and form a complex)
then specific antibody is added to bind to the antigen: this antibody is linked to an enzyme that will change color if the antigen antibody complex is formed, otherwise it’s washed away

22
Q

reading color change in a indirect ELISA

A

lighter color = less antibody in sample

23
Q

are indirect ELISA tests more sensitive or specific?

A

increased sensitivity because there is increased antibody in the patient sample which means there is more epitope availability for secondary antibody

24
Q

indirect ELISA test result interpretation

A

in the example of FIV:
< 0.300 = negative, 0.300-0.499 = indeterminate (retest), > 0.500 = positive

25
Q

how do you perform a sandwich ELISA test?

A
  1. capture antibody coats the wells
  2. antigen to be tested is added and will bind to capture antibody (antigen in patient sample needs epitopes for both capture and detection antibody)
26
Q

which disease is the sandwich ELISA test typically used for?

A

common for feline leukemia

27
Q

is the sandwich ELISA test more specific or sensitive?

A

highly specific and has increased sensitivity

28
Q

competition/inhibition blocking ELISA test (bELISA)

A
  1. wells are coated with a capture antibody
  2. antigen is added which forms complex
    if the sample has increased antibodies to the antigen, it will compete with the detection antibody and will wash away so there will be significant reduction in signal
29
Q

is the bELISA test more sensitive or specific?

A

increased sensitivity because we are looking for a specific antibody to antigen

30
Q

how is a bELISA test read?

A

color change is opposite: negative = darker color and positive = lighter color

31
Q

immunodiffusion test

A

Coggins test
antigen and patient sample are added to the wells on an agar plate —> both will diffuse into the agar and form a visible band of precipitation as a reaction
a nonidentity reaction will occur when the Ag-Ab complexes are different (the resulting “X” or crossed reaction indicates that 2 unrelated complexes are present)

32
Q

what are 2 other immunodiffusion tests?

A

fluorescent antibody and RIA tests

33
Q

fluorescent antibody test

A

antibodies from the patient are added to a fluorescent dye-coated antigen slide which is viewed under a fluorescent mircoscope

34
Q

RIA test

A

radioimmunoassay
determines antibody levels by attaching radioisotope to antigens and measuring radioactivity

35
Q

antibody titer

A

measurement of how many antibodies an animal has produced against a specific antigen
done by making serial dilutions of the sample

36
Q

antibody titer results

A

low titer = previous exposure
high titer = active infection
booster vaccine will elicit a secondary response (IgG) which will give you a higher titer result

37
Q

seroconversion

A

negative antibody converts to positive antibody titer or low antibody titer converts to high level (via vaccine or exposure)

38
Q

seroreversion

A

antibody decay

39
Q

seroprevalence

A

proportion of seropositive animals in a population

Clinical Pathology II (15 decks)

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