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Pathology > Lecture 11: Immunologic tests > Flashcards

Lecture 11: Immunologic tests Flashcards

1
Q

immunologic tests

A

Useful in confirming diagnosis

Often necessary before initiating treatment or making a referral

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2
Q

indications for immunologic testing

A

Blood-typing
Auto-antibody detection
Immune deficiency testing
Pathogen detection

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3
Q

when might we need to order immunologic tests in optometric care

A 
Ocular infections
     Conjunctivitis
      Vitritis
Autoimmune disease
    Uveitis
    Dry eye
    Amaurosis fugax
Severe ocular Allergies
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4
Q

what is serology and how is it used clinically

A

Study of blood serum and other bodily fluids

Clinically: diagnostic identification of antibodies in serum

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5
Q

What is Diagnostic Immunology (Immunodiagnostics) and what 3 things are used to identify the target molecules?

A

Ag or Ab detection through the use of Ag:Ab interactions

Ag or Ab identified by:
Radiolabel
Enzyme
Fluorescent label

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6
Q

Lab-grown, Ag-specific Abs are manufactured by creation of ___

A

hybridoma

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7
Q

hybridoma

A

fast-growing cell line produced by fusing a cancer cell (myeloma) to another cell, such as an antibody-producing cell

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8
Q

3 uses of mAB

A

Immunodiagnostics
Cancer treatment
Autoimmune disease therapy

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9
Q

Naked mAbs = ___ Abs

A

independent

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10
Q

what are conjugated mABs and what are three examples

A

Abs joined with another molecule:
Chemotherapy drug
Radioactive particle
Toxin

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11
Q

5 serologic tests

A
  1. Precipitation tests
  2. Agglutination tests
  3. Complement fixation tests
  4. Viral neutralization tests
  5. Labeled antibody tests
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12
Q

precipitation test

A

Soluble target molecule forms a precipitate in test medium after binding w/ known Ag or known Ab:
Often visible to naked eye
Requires large amounts of Ag or Ab for positive test
Low sensitivity

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13
Q

2 types of precipitation tests

A

Gel immunodiffusion

Immunoelectrophoresis

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14
Q

what are steps to gel gel immunodiffusion

A

Gel immunodiffusion:

  1. Serum placed in one well
  2. Solution w/ known Ag placed in one well
  3. Precipitation line forms if Ab present is sample
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15
Q

immunoelectrophoresis

A

Tests for Ag or Ab:
Proteins propelled across gel by electrical charge
Separated proteins diffused against Abs trough
Precipitation indicates presence of target Ag

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16
Q

crossed immunoelectrophoresis

A

Proteins electrically propelled into Ab-doped gel 90 degrees from initial direction
Height of precipitation line allows for quantification of Ag present in sample

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17
Q

agglutination tests

A

Tests for virus, bacteria, and Abs:
Cross-linking of insoluble particles causes clumping
Easy to see and interpret with the unaided eye
Fast
Not very sensitive

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18
Q

titration

A

Used to determine concentration of Abs or Ag in serum

Titer is recorded as reciprocal of greatest dilution which still causes reaction: 1:1000

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19
Q

hemagglutination

A

Clumping of red blood cells after mixing of anti-RBC antibodies (type of agglutination test):
Blood typing for transfusion

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20
Q

2 more types of agglutination tests

A

Latex fixation test

Flocculation test

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21
Q

latex fixation and two things it targets

A

Ab or Ag coated latex beads clump on mixing with target

Rubella virus, and rheumatoid factor

22
Q

flocculation test and what it targets

A

Foaming reaction from known Ag and Ab-containing serum

Venereal Disease Research Laboratory (VDRL) test:
Syphilis

23
Q

what are 5 labeled assays

A
  1. Radioimmunoassay (RIA)
  2. Enzyme-linked immunosorbent assay (ELISA)
  3. Western blot
  4. Immunofluorescence
  5. Cell sorting (FACS)
24
Q

RIA

A

Competitive immunoassay
Detects hormone levels in the blood

Also tests for: SLE Abs, HBsAg, drugs in plasma
 Older and cheaper than new tests, more dangerous

25
Q

What is the RIA process

A

Known quantities of radioactive hormone mixed with antibody
Add unlabeled, “cold” hormone from the patient
Measure amount of labeled hormone displaced

26
Q

ELISA

A

Test for Ag or Ab
Fast and sensitive
In vitro disease diagnostics

27
Q

examples of in vitro disease diagnostics

A

HIV, Hep B & C, SLE: antibodies in sample
Cocaine, opiates, marijuana: drug in sample
Pregnancy, thyroid function, steroids: hormone in sample

28
Q

Direct ELISA process

A
  1. Monoclonal Ab attached to solid surface
  2. Sample solution added
  3. Ab-enzyme conjugate added
  4. Substrate added
  5. Color change indicates presence of target Ag
29
Q

lateral flow test

A

Competitive or sandwich assay:
Simple device used for at-home or in-office testing
Home pregnancy test, drug tests, HIV tests

30
Q

what is the process of movement in the lateral flow test

A

Sample migrates across a series of capillary beds:

Sample pad –> conjugate pad –> reaction stripes –> wick

31
Q

RPS Adeno Detector is a lateral flow test used to detect what 2 things in the eye?

A

Viral conjunctivitis

Adenoviral antigen

32
Q

western blot

A 
Confirmatory test
Labeled antibodies bind to protein sample or
Labeled Ag bind Ab sample (HIV testing)
VERY SPECIFIC
Used to confirm ELISA
33
Q

4 examples of when you would use western blot to confirm ELISA

A
  1. Bovine Spongiform Encephalopathy (Mad Cow Disease)
  2. Lyme disease
  3. Hepatitis B
  4. HIV
34
Q

flow cytometry

A

Automatic analysis of fluorescently tagged mAbs for target antigens
Cell surface protein expression
Intracellular protein expression
Characterizing cell populations

35
Q

Fluorescence-Activated Cell Sorting (FACS)

A

Separates cell types via fluorescence detection and electrostatic charge

36
Q

Non-Treponemal Tests for Treponema pallidum (2 types)

A

Fast & cheap

  1. Venereal Disease Research Laboratory (VDRL) test
  2. Rapid Plasma Reagin (RPR)
37
Q

Venereal Disease Research Laboratory (VDRL) test

A

Serum Ab detection via flocculation

Can be negative in latent syphilis or after treatment

38
Q

rapid plasma reagin (RPR)

A

Same Ag used as VDRL, but bound to carbon particle

Flocculation is visible w/o microscope

39
Q

Treponemal Tests for Treponema pallidum

A

Fluorescent Treponemal Antibody Absorption Test (FTA-ABS)
Expensive, confirmatory test
Tests for organism-specific Abs and organism in patient serum:
Anti-treponeme Ab
Anti-human Ab

40
Q

2 tests for inflammation

A
  1. C-reactive protein (CRP): acute inflammation

2. Erythrocyte sedimentation rate (ESR)

41
Q

Creactive protein

A

Non-specific
Acute-phase protein
Associated with complement activation

42
Q

Erythrocyte sedimentation rate

A

Non-specific
Measure of RBC sedimentation
Tests for inflammation
Acute or chronic

43
Q

___ ____ is produced by the liver during an acute inflammatory response

A

c-reactive protein

44
Q

CRP levels are associated with elevated _____ risk

A 
Disease:
Heart attack
Stroke
Severe macular degeneration
Colon cancer
Type II diabetes
45
Q

Erythrocyte sedimentation rate

A

Rate at which RBCs settle out of unclotted blood in one hour
Acute phase reactants (fibrinogen) accelerate RBC sedimentation
Results affected by age and sex

46
Q

rheumatoid factor

A

Primary serological test for rheumatoid arthritis

Detects auto-antibody that specifically binds to immunoglobulin G (IgG) in blood stream.

47
Q

__% of patients with RA are RF+

A

70

48
Q

3 other RF+ autoimmune diseases

A
  1. SLE
  2. Sjogren’s syndrome
  3. Only 5% of Juvenile Rheumatoid Arthritis (JRA) patients are RF positive
49
Q

antinuclear antibodies

A

Detects auto-antibodies that bind host DNA

50
Q

5 Other ANA+ (antinuclear antibody) autoimmune diseases

A 
80% of patients with eye involvement of JRA
30% of rheumatoid arthritis
40-70% with Sjogren’s Syndrome
60-90% scleroderma
95% with SLE
51
Q

other ANA+ conditions

A

Syphilis

Pathology (13 decks)

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