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Immunology > Serology > Flashcards

Serology Flashcards

0
Q

Serology

A

study of serum

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1
Q

Immunodiagnostics are used for…

A
  1. Detection of Ags
  2. Detection of Abs
  3. Diagnosis of immunologic diseases
  4. Measurement of blood concentrations of pharmacologic agents/hormones
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2
Q

General formula for testing for Ags

A

Sample + Abs => detectable Ag:Ab reaction (if present in sample)

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3
Q

General formula for testing for Abs

A

Sample + Ags => detectable Ag:Ab reaction (if present in sample)

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4
Q

Antiserum

A

serum from animal deliberately immunized for a particular Ag & that has Abs to react with that particular Ag

generally said to be polyclonal

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5
Q

monoclonal antibodies

A

Abs derived from a single clone of B-cells

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6
Q

Antiglobulins

A

Abs that react with immunoglobulins of a different species because those Igs make good Ags in the different species.

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7
Q

Antiglobulins use in diagnostic tests

A

used to detect Abs from a particular species

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8
Q

False Positive

A

test is positive but the animal isn’t infected with the agent

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9
Q

False Negative

A

Test is negative but the animal is infected with the agent

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10
Q

False Positive

testing for Ag

A
  • recently vaccinated for the agent (& ags still present)

- infected with an agent that cross-reacts

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11
Q

False Negative

testing for ag

A
  • low level of Ag present in specimen
    • early in infection
    • late in infection
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12
Q

False Positive

testing for ab

A
  • cross-reactive abs are being detected

- previous vaccination or infection

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13
Q

False Negative

testing for ab

A
  • low level of Ab present in specimen
    • early in infection
    • poor immune response to infection
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14
Q

Secondary Binding Assays

A

detect consequences of Ag:Ab binding

aka detect function!

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15
Q

Agglutination

A

USED TO DETECT Abs (& occasionally Ag)

  • Ag is on a particle
  • specific Abs cross-link the particles => clumping of particles
  • Ag:Ab reaction detected by clumping

can be done on a slide or in wells (can do titers)

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16
Q

Agglutination to test Ags done by..

A

latex particles coated with Abs

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17
Q

Immunoprecipitation

A

NOT an immunodiagnostic test. (basis for other tests)

  • Ag in solution
  • specific Abs cross-link soluble Ags for large precipitating complexes
  • Precipitate forms when Ag & Ab at ~equivalent [molar]
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18
Q

Agar Gel Immunodiffusion (AGID) Test

A

Detect Abs

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19
Q

Agar Gel Immunodiffusion (AGID) test

performed by…

A
  • Ags & Abs placed in wells cut into an agar gel matrix
  • Ags & Abs diffuse in agar & form precipitate when they meet at optimal proportions
  • Ag:Ab reaction detected by formation of line of precipitate (precipitin line)
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20
Q

Complement Fixation

A

detects Abs

reported as titers

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21
Q

Complement Fixation

performed by..

A
  • Ags & Abs mixed & source of complement is added
  • Ag:Ab reaction detected by indicator system (sheep RBCs [SRBC] & Ab to SRBCs)
  • no reaction in 1st step, complement can lyse SRBCs coated w/ Abs.
  • reaction in 1st step, complement is consumed & no SRBCs lysed.
22
Q

Complement Fixation results

A

positive test: Absence of SRBC lysis (complement consumed)

negative test: Presence of SRBC lysis (complement available)

23
Q

Hemagglutination Inhibition

A

clinically, detect Abs
in lab, can ID virus isolated from clinical specimens using Abs w/ known specificity

done in a plate & reported as a TITER

24
Q

Hemagglutination Inhibition mechanism

A
  • some viruses cause agglutination of RBCs of certain species
  • Ag:Ab reaction detected by inhibition of hemagglutination by virus
25
Q

Serum Neutralization (SN) Test

A

Used to detect neutralizing Abs

Results reported as Titers

26
Q 
Serum Neutralization (SN) Test mechanism
for viruses
A
  • Virus grown in tissue culture cells.

- Growth detected by change in (or death) of cells

27
Q 
Serum Neutralization (SN) Test mechansim
for toxins
A

-Toxin added to cells affected by the toxin

28
Q 
Serum Neutralization (SN) Test mechanism
general
A
  • 1st, virus or toxin incubated with Abs prior to adding it to cell indicator system
  • neutralizing Abs detected by ability to inhibit damaging effects of the virus/toxin.
29
Q

Primary Binding Assays

A

detect Ag:Ab binding DIRECTLY

aka detect BINDING

30
Q

Types of Primary Binding Assays

A

Fluorescent Antibody test, ELISA (Ag & Ab), Western Blot, Immunohistochemistry, Immunochromatography

31
Q

Types of Secondary Binding Assays

A

Agglutination, Immunoprecipitation, Agar Gel Immunodiffusion Test, Complement Fixation, Hemagglutination Inhibition, Serum Neutralization test

32
Q

Fluorescent Antibody (FA) Test

A

Direct: detects Ag
Indirect: detects Ab

results reported as titers or qualitative results

33
Q

Fluorescent Antibody Test mechanism

A 
fluorescent molecule (fluorescein) covalently bonded to Ab.
Ag:Ab reaction detected by presence of fluorescence on slide
34
Q

Direct Fluorescent Antibody Test

A

tests for presence of Ag on a slide using a labeled Ab to the Ag

35
Q

Indirect Fluorescent Antibody Test

A

tests for presence of Abs to an Ag on a slide using labeled antiglobulin reagent

36
Q

Enzyme-Linked ImmunoSorbent Assay (ELISA)

for Ags

A
  • Ab to specific infectious agent attached to surface
  • Sample being tested for Ag added & unbound substances washed away.
  • Enzyme covalently attached to 2nd Ab specific for infectious agent
  • Enzyme converts colorless substrate (chromagen) to a colored product
  • Ag:Ab reaction detected by presence of color reaction when chromagen added
37
Q

Enzyme-Linked ImmunoSorbent Assay (ELISA)

for Ab

A
  • Ag of infectious agent attached to surface
  • Sample being tested for presence of Ab to the infectious agent added
  • Unbound substances washed away
  • Enzyme is covalently attached to: (1) Ag of infectious agent (2) antiglobulin
  • Chromagen converted to colored product in enzyme’s presence
  • Ag:Ab reaction detected by presence of colored reaction
38
Q

Immunoblot, Western Blot

A

detect Abs

clinically used to demonstrate specificity of Ab for particular Ag in mix of Ags

39
Q

Immunoblot, Western Blot mechanism

A
  • Ags separated according to MW by agar gel electrophoresis then transferred to a membrane
  • Ag:Ab reaction detected by addition of enzyme-labeled Ab, followed by chromogen
40
Q

Immunohistochemisty

A

tests for Ag presence in particular location in a tissue specimen

(& occasionally Ab)

41
Q

Immunohistochemistry mechanism

A
  • Ag is present in a tissue section on a slide (biopsy)

- Ag:Ab reaction detected by addition of enzyme-labeled Ab, followed by chromogen.

42
Q

Immunochromatography

A

detects Ags in blood sample

43
Q

Immunochromatography mechanism

A
  • Filter strip prepped w/ gold-labeled Abs @ one site & another site has unlabeled Abs (both recognize Ag being tested)
  • Sample added to Gold-Ab site & Ag binds to gold-Ab if present.
  • Buffer is added to cause gold-Ab to migrate toward unlabeled Ab @ 2nd site
  • If Ag present, binds to Ab @ 2nd site, concentrating gold-Ab & making line.
44
Q

Immunochromatography results in absence of Ag

A

Gold-Ab migrates to 2nd site when buffer is added, but NO line forms.

45
Q

Complement Fixation detection method

A

activation of complement

46
Q

Hemagglutination Inhibition detection method

A

inhibition of virus’s ability to agglutinate RBCs

47
Q

Enzyme-Linked ImmunoSorbent Assay (ELISA)

A

detects Ag & Ab

48
Q

Serum Neutralization detection method

A

neutralization of virus infectivity or toxin

49
Q

Agar Gel Immunodiffusion (AGID) detection method

A

precipitation of soluble antigens

50
Q

Immunoblot, Western Blot detection method

A

Ag separated according to MW by electrophoresis.

Ag or Ab labeled w/ enzyme. enzymatic reaction causes formation of colored product.

51
Q

Immunohistochemistry (IHC) detection method

A

Ags are on glass slide (tissue section or cytology).

Ab or Ag labeled with enzyme. Enzymatic reaction causes formation of colored product.

52
Q

Immunochromatography detection method

A

Ab is labeled w/ gold particles that migrate across a filter, capture of gold-labeled Ag:Ab complex leaves a visible line on the filter.

Immunology (19 decks)

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