Preparation of Serologically Active Mycobacterial Antigens.
Rept. no. 6 (Terminal), 1 Mar 69-30 Sep 75,
GEORGE WASHINGTON UNIV WASHINGTON D C
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The antigens were all prepared from M. tuberculosis in the following manner. Tubercle bacilli are cultured as surface pellicles on Proskauerbeck medium for 6 to 8 weeks and then separated from the culture fluid by membrane filtration. The culture filtrates are concentrated by prevaporation to one-tenth their original volume. After dialysis, the A protein and polysaccharide are isolated. Bacterial extracts are used as the source of the C protein. The extracts are obtained by suspending the harvested cells in isotonic saline and subjecting them to sonication in a Branson Model 575 Sonifier. Ultracentrifugation at 144,000 X G for 100 minutes provides the final cell extract from which the C protein is isolated. The concentrated culture filtrate and the cell extract are fractionated. In studies conducted using these antigens in the serodiagnosis of simian tuberculosis, it was found that the soluble antigen flourescent antibody SAFA test proved to be more sensitive and more specific than the customary intradermal tuberculin tests.