METHOD FOR THE PREPARATION OF A CHLAMYDIAE GROUP-SPECIFIC ANTIGEN ON HELA-229 CELLS INFECTED WITH A STRAIN OF CHLAMYDIA-TRACHOMATIS FOR USE IN THE COMPLEMENT-FIXATION TEST

Abstract

A simple method for the preparation of a potent group-specific antigen on HeLa-229 [human cervical carcinoma] cells infected with MRC-1 (LB) (TRIC/GB/MRC-1 Gf) strain of C. trachomatis is outlined. HeLa-229 cells are infected (MRC-1 strain, 102 inclusion-forming units/cell) with centrifugation at 4000 g for 1 h in flat-bottomed vials. The cells are removed by brief trypsinization with 0.25% trypsin and put into 75 cm2 culture flasks (12 .times. 106 cells by flask) in BHK-21 medium supplemented with fetal bovine serum. The flasks are incubated for 5 days at 37.degree. C. The destroyed cell monolayer and the supernatant are centrifuged at 100,000 g for 1 h. The pellet is collected and resuspended in PBS [phosphate buffered saline] and subjected to ultrasonic vibration for 30 min. This antigen may be used for detection of complement-fixing antibodies in lymphogranuloma venereum and ornithosis.