The Scheme of Complement Fixation Test
Ingredients |
Test tube 1 |
Serum control 2 |
Antigen control 3 |
Patient’s serum in dilution 1:25 (ml) |
0.5 |
0.5 |
-- |
Antigen (diagnosticum) (ml) |
0.5 |
-- |
0.5 |
Complement (working dose) (ml) |
0.5 |
0.5 |
0.5 |
Isotonic solution (ml) |
-- |
0.5 |
0.5 |
Incubation at 37oC for 1 h. |
|||
Hemolytic system (ml) |
1.0 |
1.0 |
1.0 |
Incubation at 37oC for 1 h. |
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RESULTS: (hemolysis/no hemolysis) |
|
|
|
CONCLUSION: the CFT is ___________________(positive/negative). The patient’s serum contains (does not contain) specific antibodies.
|
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Picture 2. The Scheme of ELISA (the “sandwich method”)
|
Symbols and comments:
|
Table 4
The Scheme of ELISA (solid phase modification)
Ingredients |
Test |
«Positive control» |
«Negative control» |
|||||
A1 A2 |
B1 B2 |
C1 C2 |
D1 D2 |
E1 E2 |
F1 F2 |
G1 G2 |
H1 H2 |
|
1. Buffer A (ml) |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
Incubation at room temperature for 10 min. |
||||||||
2. Patient’s specimen (antigen tested) (ml) |
0.1 |
-- |
-- |
-- |
-- |
-- |
-- |
-- |
3. Standard antigen 100 μg/ml (ml) |
-- |
0 |
0 .1 |
0.1 |
0.1 |
0.1 |
0.1 pour out |
-- |
4. Reagent «negative control» (ml) |
-- |
-- |
-- |
-- |
-- |
-- |
-- |
0.1 |
Incubation at 37oC for 20 min. |
||||||||
5. Conjugate (ml) |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
Incubation at 37oC for 20 min. |
||||||||
6. Substrate (ml) |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
Incubation at room temperature for 20 min. |
||||||||
7. 50% sulfuric acid (ml) |
0.05 |
0.05 |
0.05 |
0.05 |
0.05 |
0.05 |
0.05 |
0.05 |
RESULTS: (color/no color) |
|
|
|
|
|
|
|
|
CONCLUSION: the ELISA is ________________(positive/negative). The result means, that Ag is (not) detected in the patient’s specimen. The concentration of Ag is _______ μg/ml.
|
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.1
LESSON 2 Date:
MEDICAL IMMUNOBIOLOGICAL PREPARATIONS
Table 5