Detection of Mycoplasma mycoides subspecies mycoides by monoclonal antibody-based sandwich ELISA

Abstract

Monoclonal antibodies (MAbs) were produced from a mouse immunised with Mycoplasma mycoides subsp. mycoides small colony (MmmSC) antigen and their use to detect and differentiate strains within the Mycoplasma mycoides cluster investigated in an antigen capture ELISA format. The MAbs produced could not distinguish between MmmSC and M. mycoides subsp. mycoides large colony (MmmLC) strains. However, the sandwich ELISAs developed were able to specifically distinguish these two biotypes from the other four members of the M. mycoides cluster, and from all other mycoplasma or bacteria species examined. The most sensitive application of the test was a combination of enrichment and capture by overnight or 48-h incubations of samples inoculated into mycoplasma broth in antibody-coated microtiter wells.