1. Exp Ⅱ Agglutination reaction and Antibody preparation (2)

2. Test 1 Agglutination reaction

3. 1.Principle: Tube agglutination reaction Ag-Ab reaction Agglutination reaction Slide agglutination reaction QualitativeSemi-Quantitative Test 1 Agglutination reaction

4. 1.Principle: Test 1 Agglutination reaction 1) Slide agglutination reaction: Known Ag + Unknown Ab Agglutination turbid ? 2) Tube agglutination reaction: Quantitative Ag + Serially diluted Ab Different degree of agglutination YES NO Determine the titer of Ab

5. 1.Principle: What is the titer of Ab? The titer is customarily reported as the reciprocal of the highest dilution of Ab that causes an obvious agglutination. Test 1 Agglutination reaction

6. 2. Experiments Ⅰ Tube agglutination test Ⅱ Slide agglutination test Test 1 Agglutination reaction

7. Ⅰ Tube agglutination test 1. Material: Inactivated typhiod bacteria (7×10 8 /ml), anti-serum 2. Method: 0.50.50.50.50.5 0.5 1:201:40 1:801:1601:320 0.5 Discard 12345 0.1 6 0.9 N.S Anti-H Serum Control 0.5ml ① ② ③ ④ Inactivated typhiod bacteria 0.5ml0.5ml0.5ml0.5ml0.5ml0.5ml Dilution ratio of Ab Shake 56 ℃, 1~2h

8. 3. Attentions  Sign the tube  Use the dropper tube correctly  Mix thoroughly  T: 56 ℃  Record the time  Take the tube carefully Ⅰ Tube agglutination test

9. 4. Anticipated results + Very small agglutination turbid -No agglutinationSame as control ++ Small and depositing partly Obvious turbid +++ Large and depositing mostly Slightly turbid ++++ Large and depositing Clear Degree of agglutination SedimentSupernantant Ⅰ Tube agglutination test

10. Sediment Agglutination degree The highest dilution ratio Titer of the anti-serum

11. Ⅱ Slide agglutination test 1. Material: Staphylococci Inoculation loop 2. Method: 12 Anti serum1 a. Slide b. Bacteria c. Reaction Mix bacteria with the serum d. observation Anti serum2

12. 3. Anticipated results:+- Ⅱ Slide agglutination test

13. 4. Attentions:  Sterile operation Ⅱ Slide agglutination test

14. Test 2 Secondary immunization

15. 2. Methods: Uncoagulated SRBC Add 3ml N.S and mix 2000rpm, 5min Discard the supernant Make 20% 、 25% 、 30% 、 40% SRBC solution Take 0.1ml to immunize the mice subcutaneously Add 3ml N.S and mix