Control of Infectious Synovitis. 12. Preparation of an Agglutination Test Antigen

Abstract

Lecce (12) found infectious-synovitis-derived pleuropneumonia-like organisms (ISD-PPLO) colonies growing as satellites to Micrococcus colonies. This was confirmed by Chalquest and Fabricant (5), who grew the organism in Difco PPLO broth enriched with 10% swine serum, 0.1% diphosphopyridine nucleotide (DPN), and 0.1% 1-cysteine (free base). Chalquest (4) later found the optimum level of cysteine and DPN to be 0.01%, and that cysteine hydrochloride could be used in place of the free base. Growth was better on agar containing 0.5% soluble starch and 0.05% trypticase (BBL). The best maintenance medium was SD medium, which was made by incorporating 0.01% DPN in Fabricant's SA medium (7). Chalquest and Halfhill (6) produced an antigen for hyperimmunization of rabbits by using a broth medium containing Difco PPLO broth without crystal violet and supplemented with the following: 10% swine serum, 0.5% soluble starch, 0.05% trypticase, 0.005% cysteine hydrochloride, and 0.01% DPN. Klieneberger-Nobel (11) found that antigen production was better with solid agar in the medium. Many slight variations in procedures and media have been used in preparing Mycoplasma antigens (1, 2, 3, 8, 9). These seem to be related to the practices of the individual laboratory. This report deals with the production of ISD-PPLO antigen from agent 1853 (14) and its use in serum-plate-, serum-tube-, and whole blood agglutination tests.