Precipitation tests

Agar immunodiffusion
Lab 5

Serological Tests

Precipitation tests
Antigen/Ab conjugation
Neutralisation tests
Agglutination tests
ELISA tests
Complement Fixation Tests (CF)

Precipitation tests
One of the properties of some antibody
classes is the ability to precipitate from
solution when combined with multivalent
antigens; such reactions can be visualized
This behavior is called precipitation

Agglutination tests
Antibodies can agglutinate multivalent
particulate antigens, such as red blood
cells or bacteria
This behavior is called agglutination.
Serological tests based on agglutination
are usually more sensitive than those
based on precipitation

Neutralisation tests
Usually used for viral or bacterial/toxin
identification
Antibody neutralises the toxin and
prevents its action (antitoxins)
Requires an indicator system e.g. lab animals,
tissue culture, etc

Antigen/Ab conjugation
Antigens (or immunoglobulins) can be
conjugated with other molecules
(radioisotopes, enzymes or fluorescent
dyes) so that antigen-antibody binding can
be detected at extremely low
concentrations.
Examples are radioimmunoassay or
enzyme-immunoassay, Fluorescent
antibody tests (FAT).
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Guide to the use of Serological Tests

Which test?
Consider :
the organism
the most accurate test (sensitivity/specificity)
quantitative vs qualitative
the legal status of the test
the material (sample) submitted

Precipitation Tests
Used for soluble antigen
When soluble antigen reacts with its specific
antibody in solution, the antigen-antibody
complex may become insoluble and precipitate
Relatively less sensitive

Optimal proportions of antigen and

antibody causes precipitation
Examples
Excess antibody or antigen no
precipitation
Ring precipitation test

Lancefield grouping
Agar gel immunodiffusion (AGID)
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Ring precipitation test

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Lancefield grouping

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Agar Gel Immunodiffusion

Agar gel in plates or on microscope slides
Make wells in agar for both antigen and
antibody
Two reactants diffuse into agar and form
immunoprecipitates where optimal
proportions occur

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Reaction of identity

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Reaction of partial identity

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Reaction of non-identity

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Radial Immunodiffusion
Antiserum

Antigen

Decreasing Antigen
Concentration

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Log Ag
conc.
Diameter of the precipitation ring

Prepare a standard curve

Allows exact quantification of antigen
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