A filter immunobinding technique for the rapid detection and simultaneous identification of avian and bovine mycoplasmas.
A filter immunobinding (FIB) method was developed for detecting and identifying mycoplasmas. Nine avian and 9 bovine mycoplasma strains were propagated in broth media and were diluted and immobilized on a nitrocellulose membrane as antigens. Non-specific FIB reactions were easily eliminated by absorbing rabbit hyperimmune serum in the broth. Absorbed rabbit hyperimmune sera showed clear species-specificity with mycoplasma antigens by the FIB. These specific reactions were in agreement with biochemical and growth inhibition tests for Mycoplasma bovirhinis, M. bovis, M. columbinum, M. columborale, M. gallisepticum and M. synoviae. Some bovine mycoplasma strains, which could not be identified by growth inhibition test, because of their strong production of film and spots on the agar, specifically reacted with absorbed rabbit hyperimmune sera against M. bovis in the FIB. The detection limit of mycoplasmas by this method was about 104-105 c.f.u./ml, which is lower than that of colony determination on agar. It is suggested that FIB may be a useful technique for rapid detection and identification of mycoplasmas.