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Serology Flashcards

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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..

Study These Flashcards
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

Study These Flashcards
A

positive test: Absence of SRBC lysis (complement consumed)

negative test: Presence of SRBC lysis (complement available)

23
Q

Hemagglutination Inhibition

Study These Flashcards
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
Hemagglutination Inhibition mechanism
- some viruses cause agglutination of RBCs of certain species - Ag:Ab reaction detected by inhibition of hemagglutination by virus
25
Serum Neutralization (SN) Test
Used to detect neutralizing Abs | Results reported as Titers
26
``` Serum Neutralization (SN) Test mechanism for viruses ```
- Virus grown in tissue culture cells. | - Growth detected by change in (or death) of cells
27
``` Serum Neutralization (SN) Test mechansim for toxins ```
-Toxin added to cells affected by the toxin
28
``` Serum Neutralization (SN) Test mechanism general ```
- 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
Primary Binding Assays
detect Ag:Ab binding DIRECTLY aka detect BINDING
30
Types of Primary Binding Assays
Fluorescent Antibody test, ELISA (Ag & Ab), Western Blot, Immunohistochemistry, Immunochromatography
31
Types of Secondary Binding Assays
Agglutination, Immunoprecipitation, Agar Gel Immunodiffusion Test, Complement Fixation, Hemagglutination Inhibition, Serum Neutralization test
32
Fluorescent Antibody (FA) Test
Direct: detects Ag Indirect: detects Ab results reported as titers or qualitative results
33
Fluorescent Antibody Test mechanism
``` fluorescent molecule (fluorescein) covalently bonded to Ab. Ag:Ab reaction detected by presence of fluorescence on slide ```
34
Direct Fluorescent Antibody Test
tests for presence of Ag on a slide using a labeled Ab to the Ag
35
Indirect Fluorescent Antibody Test
tests for presence of Abs to an Ag on a slide using labeled antiglobulin reagent
36
Enzyme-Linked ImmunoSorbent Assay (ELISA) | for Ags
- 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
Enzyme-Linked ImmunoSorbent Assay (ELISA) | for Ab
- 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
Immunoblot, Western Blot
detect Abs clinically used to demonstrate specificity of Ab for particular Ag in mix of Ags
39
Immunoblot, Western Blot mechanism
- 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
Immunohistochemisty
tests for Ag presence in particular location in a tissue specimen (& occasionally Ab)
41
Immunohistochemistry mechanism
- 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
Immunochromatography
detects Ags in blood sample
43
Immunochromatography mechanism
- 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
Immunochromatography results in absence of Ag
Gold-Ab migrates to 2nd site when buffer is added, but NO line forms.
45
Complement Fixation detection method
activation of complement
46
Hemagglutination Inhibition detection method
inhibition of virus's ability to agglutinate RBCs
47
Enzyme-Linked ImmunoSorbent Assay (ELISA)
detects Ag & Ab
48
Serum Neutralization detection method
neutralization of virus infectivity or toxin
49
Agar Gel Immunodiffusion (AGID) detection method
precipitation of soluble antigens
50
Immunoblot, Western Blot detection method
Ag separated according to MW by electrophoresis. | Ag or Ab labeled w/ enzyme. enzymatic reaction causes formation of colored product.
51
Immunohistochemistry (IHC) detection method
Ags are on glass slide (tissue section or cytology). | Ab or Ag labeled with enzyme. Enzymatic reaction causes formation of colored product.
52
Immunochromatography detection method
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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