Immunobinding Assay for the Speciation of Avian Mycoplasmas Adapted for Use with a 96-Well Filtration Manifold

Abstract

An immunobinding assay capable of distinguishing among Mycoplasma synoviae, M. gallisepticum, M. gallopavonis, and M. meleagridis was developed. A low-protein-binding membrane filter served as the solid support, and a 96-well microsample filtration manifold was used. The assay detected 50 ng of mycoplasmal protein, or approximately 3 x 10(4) colony-forming units, from pure or mixed cultures. No cross-reactions were observed among the mycoplasma species tested. The assay was inexpensive, easy to interpret, and required approximately 3 hr to perform. The 96-well format allowed screening of numerous cultures and use of several antisera in a single assay.