Identification of mycoplasma isolates by ELISA

An antigen-based enzyme-linked immunosorbent assay (ELISA) was developed for the rapid identification of Mycoplasma gallisepticum and Mycoplasma synoviae. Two trials were conducted to compare the ELISA with the conventional fluorescent antibody technique. In trial 1, broilers were inoculated with M. gallisepticum F, M. gallisepticum S6, and M. synoviae. In trial 2, hens were inoculated with M. gallisepticum F, M. synoviae, and a combination of the two. Swab isolates were obtained from the choanal cleft and cultures were grown in broth media with a pH indicator. The organisms were bound to 96-well plates for the ELISA or cultured on agar for the fluorescent antibody test. Monoclonal antibodies directed to M. gallisepticum F, M. gallisepticum S6, and M. synoviae were reacted with the ELISA aliquots. The ELISA test identified the mycoplasmas in these samples and was completed within 8 hr but did not identify as many isolates as the fluorescent antibody procedure. The principal advantage of this ELISA is the ability to identify the strain of mycoplasma.