Serology

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Serology

• Eva L. Dizon, M.D.
• Microbiology
• Our Lady of Fatima University

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Serology
Antigen Antibody
Immune Complex
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Serology

• Test that depends on the Formation of Immune
  Complex
• Nephelometry
• Immunodiffusion
• Complement fixation
• Cryoglobulins

• Test in which Ag Ab is fixed to a solid surface
  
• Agglutination
• Elisa
• Electrophoresis

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Immunodiffusion
A A A
B B B
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Present in reduce concentration
Zone of equivalence precipitation
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Contaminant present in antigen
I I
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Ouchterlony

• Double diffusion
• Characterize relationship between 2 antigen
• If antigen are in the same precipitation line
  ,they fail to cross

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Ouchterlony
Ag 1
Ag 2
Ag 4
Ab
Limitations Insensitive Needs large unit of Ab
and Ag Resolve by CIE placing agar in electric
field
Ag 3
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Nephelometry

• IC in solution is monitored by spectrometry
• Formation of IC is measured in a photoelectric
  cell as optical density

Add increasing mount of antigen
Ag
Ag
Ag
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Complement fixation test
IC is fixed and consume complement
1. IC (ag ab) complement 2. IC C RBC------
no hemolysis (positive)
Ag ____ C RBC -------- hemolysis (negative)
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Cryoglobulins
This are serum globulin that precipitate at T
less than 37C To detect the presence in the
blood ---- serum Samples are incubated in 4C for
several hours ----- note for precipitation
reaction Precipitated proteins isolated by
centrifugation then assayed for IG by
nephelometry,electrophoresis
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Ag and Ab fixed to a solid surface
Agglutination test for the presence of
antibody Indirect (Passive)
Particle RBC
Ag
Ag Ag
Ab
Ag
Particle agglutination
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Neutralization test
Hemmagglutination inhibition test
Virus RBC -------- Agglutinate Virus
Ab RBC -------- No Agglutination
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ASO titer
Streptolysin O (Ag) Serum of patient
Agglutination
If Ptx had Strep pyogenes infection Ab against
Streptolysin O is present
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ELISA
Direct look for antigen Slide with Known ab
------ put serum ?Ag -------wash--------add
enzyme labeled antibody--------wash--------look
under spectrophotometer
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Elisa
Indirect look for antibody Slide with antigen
--------? Ab --------wash--------- antihuman
globulin -----------wash-------- look under
spectrophotometer
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Fluoresence test
Direct- looks for antigen 1. Slide put an
antigen 2. Add fluorescein labeled antibody 3.
Examine under the UV microscope
Indirect- antibody Slide with known
antigen Wash Add fluorescein labeled antihuman Ab
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