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ExpⅡ Agglutination reaction and Antibody preparation (2)

ExpⅡ Agglutination reaction and Antibody preparation (2). Test 1 Agglutination reaction. Test 1 Agglutination reaction. 1.Principle:. Tube agglutination reaction. Ag-Ab reaction. Agglutination reaction. Slide agglutination reaction. Qualitative. Semi-Quantitative.

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ExpⅡ Agglutination reaction and Antibody preparation (2)

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  1. ExpⅡ Agglutination reaction and Antibody preparation (2)

  2. Test 1 Agglutination reaction

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

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

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

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

  7. ⅠTube agglutination test • Material: • Inactivated typhiod bacteria (7×108/ml), anti-serum Discard 2. Method: 0.5 1 2 3 4 5 6 ② Anti-H Serum 0.1 0.5 0.5 0.5 0.5 ① 0.9 0.5ml 0.5ml 0.5ml 0.5ml N.S 0.5ml 0.5 0.5 0.5 0.5 0.5 Inactivated typhiod bacteria ③ 0.5ml 0.5ml 0.5ml 0.5ml 0.5ml 0.5ml Control 1:80 1:160 1:320 1:20 1:40 Dilution ratio of Ab Shake ④ 56℃, 1~2h

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

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

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

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

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

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

  14. Test 2 Secondary immunization

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

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