An enzyme-linked immunosorbent assay for detection of avian mycoplasmas in culture.

An antigen-capture enzyme-linked immunosorbent assay (ELISA), using monoclonal antibody (Myc-9) against Mycoplasma pneumoniae, was developed for the detection of M. gallisepticum, M. synoviae, M. meleagridis, M. iowae, M. anatis, M. iners, and M. gallinarum antigens in cultures. ELISA plates were coated with Myc-9 and reacted with the different avian mycoplasma antigens, which were detected with adsorbed polyclonal anti-mycoplasma serum followed by anti-rabbit enzyme-labeled conjugate antiserum. Nonspecific reactions and cross-reactivity were eliminated by adsorbing polyclonal antisera with a pool of heterologous antigens, and a high degree of sensitivity was obtained by both antigen enrichment and treatment with the detergent N-octyl glucoside. Under these conditions, the test detected less than 10 colony-forming units/ml. This antigen-capture ELISA appears to be a highly specific, sensitive, reproducible tool and efficient in the diagnosis of mycoplasma infection.