Identification of ruminant mycoplasmas by dot immunobinding on membrane filtration (MF dot)

Abstract

A simple method of species identification of ruminant mycoplasmas by dot immunobinding (MF dot) is described. Mycoplasmas from broth cultures were directly trapped onto the surface of low protein-binding affinity membranes by vacuum filtration. Specific polyclonal anti-mycoplasma sera were then applied. Mycoplasma-antibody complexes were then revealed by an enzyme conjugated anti-immunoglobulin. A positive reaction was indicated by the development of a color reaction when substrate was added. The method was tested on 22 type-strains representative of species, subspecies, types or yet unnamed serogroups of mycoplasmas isolated from ruminants. MF dot presented a specificity close to that which could be expected from the reference serological tests. The advantages of the MF dot over the other tests are utility, rapidity, ready standardization and it allows many samples to be treated simultaneously.