THE FIRST SECHENOV MOSCOW STATE MEDICAL UNIVERSITY. Medical Faculty
General microbiology (continued from spring semester)
SECTION 3: INFECTION and IMMUNITY (continued from spring semester)
LESSON 1 Date:
ANTIGEN - ANTIBODY REACTIONS: Lysis Tests, Complement-Fixation Test, IF-test, ELISA
Table 1
The Scheme of the Immune Hemolysis Test
Ingredients |
Test 1 |
Control 2 |
Control 3 |
Control 4 |
Control 5 |
Rabbit's hemolytic antiserum (ml) |
0.5 |
0.5 |
-- |
0.5 |
-- |
Sheep erythrocytes suspension (ml) |
0.5 |
0.5 |
0.5 |
-- |
0.5 |
Complement in dilution 1:10 (ml) |
0.5 |
-- |
0.5 |
0.5 |
-- |
Dog's erythrocytes suspension (ml) |
-- |
-- |
-- |
0.5 |
-- |
Isotonic solution (ml) |
-- |
0.5 |
0.5 |
-- |
1.0 |
Total volume of the mixture (ml) |
1.5 |
1.5 |
1.5 |
1.5 |
1.5 |
Incubation at 37oC for 1h. |
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RESULTS: (hemolysis/no hemolysis) |
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CONCLUSION: the immune hemolysis test is ____________________ (positive/negative).
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Table 2
The Scheme of the Complement Titration Test
Ingredients |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
Control 9 |
Control 10 |
Complement in 1:10 dilution (ml) |
0.15 |
0.2 |
0.25 |
0.3 |
0.35 |
0.4 |
0.45 |
0.5 |
0.5 |
-- |
Isotonic solution (ml) |
1.35 |
1.3 |
1.25 |
1.2 |
1.15 |
1.1 |
1.05 |
1.0 |
1.5 |
1.5 |
Hemolytic system (ml) |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
0.5 |
1.0 |
Incubation at 37oC for 30 min. |
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RESULTS: (hemolysis/no hemolysis) |
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CONCLUSION: the titer of complement is _______ ml; the working dose of complement is _________ ml.
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Picture 1. The Scheme of Complement Fixation Test (CFT)
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Symbols and comments:
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Table 3