Immunobinding assay for the speciation of avian mycoplasmas adapted for use with a 96-well filtration manifold

An immunobinding assay capable of distinguishing among Mycoplasma synoviae, M. gallisepticum, M. gallopavonis, and M. meleagridis was developed. A low-protein-binding 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