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Lab techniques 2 > Immunology: Reference Lab Tests > Flashcards

Immunology: Reference Lab Tests Flashcards

1
Q

What are the most common reference lab tests?

A

Coomb’s testing
Fluorescent Antibody Testing
Antibody titers
Polymerase Chain Reaction (PCR)

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2
Q

What is a very specific test for IMHA?

A

Coombs’ test

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3
Q

What does the Coombs’ test test for?

A

The presence of autoantibodies

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4
Q

What are autoantibodies

A

Antibodies against the body

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5
Q

What does a Direct Coombs’ test test for?

A

Detects antibodies that attack RBCs

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6
Q

What is Coombs’ test based on?

A

The level of agglutination present

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7
Q

What diseases does Immunodiffusion detect?

A

Equine Infectious anemia (retrovirus)

Johne disease

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8
Q

What is Johne (Yoh-nee) disease?

A

Ruminant GI disease caused by mycobacterium

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9
Q

What is present in the immunodiffusion test kit?

A

The antigen

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10
Q

What is placed in the in separate wells on an agar plate in an immunodiffusion test?

A

The antigen and patient serum

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11
Q

What diffue through the agar plate and form a band of precipitation where they meet in the Immunodiffusion test?

A

The antigen and patient serum

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12
Q

What does it mean if no band is present in an immunodiffusion test?

A

No antibody present

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13
Q

What is mainly used in research and diagnostic laboratories?

A

Radioimmunoassay

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14
Q

What is the difference between radioimmunoassay and CELISA tests?

A

Radioimmunoassy uses a radioisotope instead of an enzyme

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15
Q

How is Radioimmunoassay similar to CELISA?

A

It gives the amount of antigen present

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16
Q

What is not common in veterinary practice but available in reference laboratories and used to verify a tentative diagnosis?

A

Indirect Fluorescent Antibody Testing

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17
Q

Why do we run an IFA

A

To identify a specific antibody in a sample

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18
Q

How does the direct antibody fluorescent antibody test work?

A

The patient sample is added to a fluorescent dye-conjugated antigen and if antigen/antibody complexes form (the disease is present) the sample will fluoresce

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19
Q

What does an indirect Coombs test test for?

A

Detects antibodies that are already attached to red blood cells

20
Q

What does a positive Coombs test mean?

A

Immune-mediated hemolytic disease

21
Q

What kind of sample does the Radioimmunoassay test need?

A

Serum

22
Q

In Radioimmunoassay testing, how do patient antigen and labeled antigen affect each other?

A

With increasing amounts of patient antigen, more labeled antigen is displaced from the antibody

23
Q

In Radioimmunoassay testing what is compared with a standard curve to determine the concentration of the antigen?

A

Patient’s serum

24
Q

What kind of disease do we run the IFA to rule out?

A

Diseases that have no other tests to detect them

25
Q

What is a reason we run antibody titers?

A

To figure out if an animal has a current infection or if it has prior exposure to a specific antigen

26
Q

What do antibody titers determine the need of?

A

Revaccination

27
Q

How do you run an antibody titer?

A

WIth serial dilutions of each sample and examination of each dilution for the presence of antibodies

28
Q

What does a high titer mean?

A

Indicates an active infection

29
Q

What does a low titer mean?

A

Usually indicates a previous exposure to the specific antigen

30
Q

What are molecular diagnostic testing based on?

A

The testing of DNA or RNA

31
Q

What are DNA tests used for

A

Identify cancers
Detect genetic defects
Pedigrees
Bacterial contaminants in foods

32
Q

What are the advantages of molecular diagnostics?

A

Increased sensitivity and specificity
Small samples with faster results
Factors that influence other procedures are not as crucial

33
Q

What are the disadvantages of molecular diagnostics?

A

Contamination gives false positives
High levels of expertise to run tests
Need more than one room to perform
High costs

34
Q

Polymerace Chain Reaction (PCR)

A

Detect DNA segments

Widely used

35
Q

Reverse transcriptase polymerase chain reaction

A

Similar to PCR but tests single-stranded RNA

36
Q

What must be converted to a double-stranced DNA before the PCR process can continue?

A

The single-stranded RNA

37
Q

What are the benefits of running a Real-time Polymerase Chain Reaction?

A

Decreases the risk of contamination
More easily automated
Faster and easier to run
Fluorescence probe attaches to DNA segments

38
Q

What else is the PCR called?

A

The amplification assay

39
Q

Why is the PCR called an Amplification Assay?

A

Because a small amount of DNA segment detected in the sample is amplified to run the test better and to determine the results

40
Q

What are the three steps of the amplification assay?

A

Denaturation
Annealing
Extension

41
Q

How often is this process repeated?

A

25 to 30 times to gain enough DNA

42
Q

Denaturation

A

Heated to break apart double stranded DNA

Each strand serves as a template for new nucleotides

43
Q

Annealing

A

Temperature is lowered

Causes primers to bind to separate strands

44
Q

Extension

A

Temperature raised again adn the Taq DNA polymerase causes new DNA segments to be produced

45
Q

What is Taq DNA polymerase

A

The enzyme can read DNA code and assemble the nucleotide base to form new complementary strands

Lab techniques 2 (38 decks)

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