Mycoplasma detection in cell culture by concomitant use of bisbenzamide and fluoresceinated antibody

Conditions are presented for application of both bisbenzamide (Hoechst 33258) stain and a specific fluoresceinated anti-Mycoplasma hyorhinis IgG to a single cell culture preparation. This allows the same field on a slide to be viewed for presumptive diagnosis of any cell culture contaminant mycoplasma by bisbenzamide staining and for definitive diagnosis of M. hyorhinis strains using fluoresceinated antibody. The use of this method plus a cultural procedure will permit identification of the "noncultivable" M. hyorhinis strain DBS 1050.