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Basic Pathology > Immunologic Tests > Flashcards

Immunologic Tests Flashcards

1
Q

immunologic tests

A
  • confirm diagnosis

- often necessary before treatment or referral

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2
Q

immunologic tests indications

A
  • indicators of infection and/or inflammation
  • pathogen detection
  • auto-antibody detection
  • blood and tissue typing
  • immune deficiency testing
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3
Q

indications for immunologic tests in optometric care

A
  • ocular infections (conjunctivitis, viritis)
  • autoimmune disease (uveitis, keratoconjunctivitis sicca, amuarosis fugax (sudden loss of vision))
  • ocular allergies
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4
Q

available in-office immunologic tests in optometric care

A
  • adenoviral conjunctivitis
  • Sjorgren’s syndrome
  • non-specific dry eye
  • environmental allergies
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5
Q

immunologic tests categories

A
  • serology

- diagnostic immunology (immunodiagnostics)

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6
Q

serology

A
  • study of blood serum and other bodily fluids

- clinically: diagnostic identification of antibodies, enzymes, or minerals

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7
Q

diagnostic immunology (immunodiagnostics)

A
  • antigen or antibody detection through the use of Ag:Ab interactions
  • Ag or Ab identified by radiolabel, enzyme, or fluorescent label
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8
Q

monoclonal antibodies (mAb)

A

-Ag-specific Abs lab-grown via hybridoma

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9
Q

uses of monoclonal antibodies

A
  • immunodiagnostics
  • cancer treatment
  • autoimmune disease therapy
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10
Q

naked mAbs

A

independent Abs

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11
Q

conjugated mAbs

A

Abs joined with another molecule

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12
Q

serologic tests

A
  • precipitation tests
  • agglutination tests
  • labeled antibody tests
  • complement fixation tests
  • viral neutralization tests
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13
Q

complement fixation tests

A
  • mix sample serum with test solution
  • then add to solution containing complement
  • if Abs are present in sample serum, those Abs will bind the Ags, preventing Ag from activating complement
  • presence of Abs in serum prevent complement activation
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14
Q

viral neutralization tests

A
  • put patient sample and virus in an egg
  • presence of Abs in serum prevent virus growth
  • viral growth will occur if the patient sample does not have the Abs
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15
Q

precipitation tests:

tests for _____

A
  • Abs
  • viruses
  • bacteria
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16
Q

precipitation tests:

characteristics

A
  • soluble target molecule and known Ag or Ab
  • precipitate (immune complexes) often visible to naked eye
  • requires large amounts of Ag or Ab for positive test
  • low sensitivity
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17
Q

precipitation tests:

types of tests

A
  • gel immunodiffusion

- immunoelectrophoresis

18
Q

agglutination tests:

tests for ____

A
  • Abs
  • viruses
  • bacteria
19
Q

agglutination tests:

characteristics

A
  • target may be bound to a carrier protein (increases visibility)
  • cross-linking causes clumping
  • easy to see and interpret
  • fast
  • can be very sensitive
20
Q

hemagglutination

A
  • clumping of red blood cells after mixing of anti-RBC antibodies
  • blood typing for transfusion
21
Q

titration and agglutination

A
  • concentration of Abs or Ag in serum
  • titer = reciprocal of greatest dilution with reaction
  • e.g., determining IgG titer after vaccination
22
Q

agglutination tests for Treponema pallidum (syphilis pathogen)

A
  • fast and cheap but not very specific
  • Venereal Disease Research Laboratory (VDRL) test
  • Rapid Plasma Reagin (RPR)
23
Q

Venereal Disease Research Laboratory (VDRL) test

A
  • serum Ab detection via flocculation

- can be negative in latent syphilis or after treatment

24
Q

Rapid Plasma Reagin (RPR)

A
  • same Ag used as VDRL, but bound to carbon particle

- flocculation is visible without a microscope

25
Q

labelled assays

A
  • radioimmunoassay (RIA)
  • enzyme-linked immunosorbent assay (ELISA)
  • immunofluorescence
  • western blot
  • flow cytometry
26
Q

radioimmunoassay (RIA)

A
  • competitive immunoassay
  • detects hormone levels (T3, T4)
  • also tests for: SLE Abs, HBsAg, drugs in plasma
27
Q

enzyme-linked immunosorbent assay (ELISA)

A
  • test for Ag or Ab - with visible color change
  • fast and sensitive
  • in vitro disease diagnostics: Abs in sample (HIV, SLE, Grave’s, myasthenia gravis); drug in sample (cocaine, opiates, marijuana); hormone in sample (pregnancy, thyroid function, steroids)
28
Q

sandwich ELISA

A

1) monoclonal Ab attached to solid surface
2) sample solution added
3) Ab-enzyme conjugate added
4) substrate added
5) target Ag causes color change
6) can titrate to find concentration in sample

29
Q

rheumatoid factor

A
  • auto-Abs that binds Fc region of IgG

- detected via ELISA

30
Q

RF+ autoimmune diseases

A
  • 70% of patients with RA are RF+
  • SLE
  • Sjogren’s syndrome
  • only 5% of Juvenile Rheumatoid Arthritis (JRA)
31
Q

lateral flow immunoassay

A
  • competitive or sandwich assay
  • qualitative, not quantitative
  • simple device used for at-home or in-office testing (home pregnancy tests, drug tests, HIV tests)
  • sample migrates across a series of capillary beds
  • sample pad –> conjugate pad –> reaction stripes –> wick
32
Q

lateral flow assays for ocular disease

A
  • RPS Adeno Detector (viral conjunctivitis, adenoviral antigen)
  • RPS Inflammatory (dry eye, matrix metalloproteinase-9)
33
Q

immunofluorescence and Antinuclear Antibodies (ANA)

A
  • auto-Abs that bind host DNA, RNA, etc.

- indirect fluorescent antibody (IFA): mix serum sample with cells fixed to slide, add fluorescently labeled Abs

34
Q

ANA+ autoimmune diseases

A
  • 80% of patients with eye involvement of JRA
  • 95% with SLE
  • 30% of rheumatoid arthritis
  • 40-70% with Sjogren’s syndrome
  • 60-90% scleroderma
35
Q

Western blot

A
  • confirmatory test
  • labeled antibodies bind to protein sample or labeled Ag bind Ab sample (HIV testing)
  • very sensitive and very specific
  • used to confirm ELISA (bovine spongiform encephalopathy, Lyme disease, hepatitis B, HIV)
36
Q

automated analysis by flow cytometry

A
  • automatic analysis
  • fluorescently tagged mAbs
  • cell surface protein expression
  • intracellular protein expression
  • characterizing cell populations
37
Q

treponemal test for treponema pallidum

A
  • Fluorescent Treponemal Antibody Absorption Test (FTA-ABS)
  • expensive, confirmatory test (would use this if someone had a positive RPR)
  • looking for organism-specific Abs (IgG and IgM)
  • add sample serum to slide with fixed pathogen
  • add fluorescently tagged anti-human mAb
  • look for anti-treponeme Abs on surface of treponemes
38
Q

blood tests for inflammation

A
  • C-reactive protein (CRP): acute inflammation

- Erythrocyte sedimentation rate (ESR): acute and chronic inflammation

39
Q

C-reactive protein (CRP)

A
  • non-specific (tells you someone has inflammation but not why)
  • produced by the liver during an acute inflammatory response
  • associated with complement activation
  • CRP levels are associated with elevated disease risk (heart attack, stroke, severe macular degeneration, colon cancer, type II diabetes)
  • detected via various immunodiagnostic assays (ELISA)
40
Q

Erythrocyte sedimentation rate (ESR)

A
  • non-specific
  • measure of RBC sedimentation
  • rate at which RBCs settle out of blood in one hour
  • treated to prevent coagulation
  • acute phase reactants (fibrinogen) accelerate RBC sedimentation
  • results affected by age and sex
  • good for monitoring disease

Basic Pathology (21 decks)

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