[Standardization of the complement fixation test (CFT) in brucellosis. III. The hemolytic system].
The purpose of this paper was to developed simple and accurate methods for standardization of the particular components of a hemolytic system and to choose optimal amounts of these components in CFT. On the basis of comparative studies and the data from literature the following procedure was chosen: 1. Sheep's blood was conserved with Alsever liquid and used after 5 days of stabilization. A 2% suspension of erythrocytes was standardized by centrifuging in calibrated test tubes at 1000 r for 10 min and spectrophotometrically, assuming the following criterion: the suspension of erythrocytes dissolved in distilled water at 1:10 ratio should show 0.500 extinction (E541), which corresponds to 5 X 10(8) erythrocytes in 1 cm3. 2. Lyophilized hemolysin was dissolved, conserved with glycerol and used after 5 days of stabilization. Hemolysin was titrated with 100% hemolysis in the presence of the excess of complement. The suspension of erythrocytes was sensitized with 5 units of hemolysin directly before using it. 3. Lyophilized complement (C) was dissolved, conserved with boric acid +K2SO4 containing liquid and used after 5 days of stabilization. Titration of C was done by 50% hemolysis; 3C'H50 was used in CFT. 4. A barbital buffer was used as diluent at pH 7.3-7.4, alternatively 0.85% NaCl with an addition of Ca and Mg at optimal concentration. The hemolytic system was incubated at 37 degrees C for 30 min.
Królak
M
M
Pracownia Badania Brucelozy Instytutu Weterynarii w Puławach z siedziba w Gdańsku.
Błaszczyk
B
B
D000818:Animals; D002007:Brucellosis, Bovine / Q000175:diagnosis*; D002417:Cattle; D003168:Complement Fixation Tests / Q000662:veterinary*; D005260:Female; D006460:Hemolysin Proteins; D006461:Hemolysis; D009996:Osmotic Fragility; D012756:Sheep
D006460:Hemolysin Proteins
Standaryzacja odczynu wiazania dopełniacza (OWD) przy brucelozie. III. Układ hemolityczny.