Improvement of PCR primers for detection of Mycoplasma mycoides subsp. mycoides, the causative agent of contagious bovine pleuropneumonia

Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease of cattle caused by Mycoplasma mycoides subsp. mycoides (Mmm). A method combining PCR and restriction enzyme analysis (PCR-REA) is described in the World Organisation for Animal Health manual to differentiate Mmm from the related species Mycoplasma mycoides subsp. capri. However, the primers in the manual did not amplify the target region from an attenuated Mmm strain. Genome sequence comparison revealed mismatches between the primers and the target sequences of Mmm strains. Newly designed primers with corrected bases successfully amplified the target region from M. mycoides strains, and REA patterns distinguished the two subspecies. These results indicate that the new primers provide reliable Mmm detection and enhance preparedness for future CBPP outbreaks.