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

2. Serology
Antigen +
Antibody
Immune Complex

3. 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

4. Increasing Antigen Concentration
Amount of Precipitine
Antigen
Excess
(Postzone)
Equivalence
Antibody
(Prozone)

5. I Immunodiffusion A B Present in reduce concentration
Zone of equivalence
+ precipitation I
Contaminant present in antigen
I I

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

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

8. 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

9. 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)

10. 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

11. Ag and Ab fixed to a solid surface
Agglutination
test for the presence of antibody
Indirect (Passive)
Particle RBC Ag Ag
+ Ab Ag
Particle agglutination

12. Neutralization test Virus + RBC -------- Agglutinate
Hemmagglutination inhibition test
Virus + RBC Agglutinate
Virus+ Ab + RBC No Agglutination

13. Agglutination ASO titer Streptolysin O (Ag) + Serum of patient
If Ptx had Strep pyogenes infection Ab against Streptolysin O is present

14. ELISA Direct – look for antigen
Slide with Known ab put serum ?Ag wash add enzyme labeled antibody wash look under spectrophotometer

15. Elisa Indirect – look for antibody
Slide with antigen ? Ab wash antihuman globulin wash look under spectrophotometer

16. 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

17. Thank you and read your Manual TOO!