A
Mycoplasma bovis PCR-based
genotyping method was developed that exploits the proximity of
insertion sequences (IS) within the
genome by using outward facing primers that selectively amplify sequences between IS elements. The method was applied to 16 field isolates of M. bovis, originating from pneumonic
lung or arthritic joints, collected from the United States (Iowa or Kansas) between 2004 and 2005. The genomic fingerprints generated 14 distinct amplification profiles consisting of 4-8 fragments
ranging in size from 200-3000 bp. Three isolates presented identical patterns and were isolated from two calves (one calf with pneumonic
lung and the other with both pneumonic
lung and
arthritic joint) from a single farm during an
outbreak and probably represent multiple infections with the same
genotype. To demonstrate the stability of IS markers for
molecular fingerprinting, 3 of the 16 field isolates were subjected to high number passage which resulted in patterns identical to the initial isolates. The results of these studies demonstrate the method can be used for simple and rapid
molecular fingerprinting and differentiating M. bovis isolates with extension to
epidemiology.