A FILTER IMMUNOBINDING TECHNIQUE FOR THE RAPID DETECTION AND SIMULTANEOUS IDENTIFICATION OF AVIAN AND BOVINE MYCOPLASMAS

A filter immunobinding (FIB) method was developed for the detection an d identification of mycoplasmas. Type strains of a total of 18 avian a nd bovine mycoplasma species propagated in broth media were diluted an d immobilized on a nitrocellulose membrane as antigens for investigati ng the specificity with rabbit hyperimmune sera. Non-specific FIB reac tions were easily eliminated by the procedure of absorbing rabbit hype rimmune sera in the broth. Absorbed rabbit hyperimmune sera exhibited clear species-specificity with mycoplasma antigens by the FIB. These s pecific reactions always agreed with the results of identification by tests of biochemical properties and growth inhibition for the isolates of M. bovirhinis, M. bovis, M. columbinum, M. columborale, M. gallise pticum and M. synoviae. Some bovine mycoplasma species, which were imp ossible to identify by growth inhibition test, because of their strong production of film and spots on the agar, specifically reacted with a bsorbed rabbit hyperimmune sera against M. bovis in the FIB. The detec tion limit of mycoplasmas by this method was about 10(4)-10(5) colony- forming units/ml, which is lower than that of colony determination on agar. The FIB seems to be a useful technique for rapid detection and s imultaneous identification of mycoplasmas.