Species identification ofMycoplasma bovisandMycoplasma agalactiaebased on theuvrC genes by PCR

The DNA repair genesuvrC fromMycoplasma bovisandMycoplasma agalactiaetype strains were cloned and their nucleotide sequences were established. These sequences were used to design polymerase chain reaction (PCR) primer pairs forM. bovisandM. agalactiae.Each primer pair amplified a 1·6 kb fragment of theuvrC gene in the respective species. The specificity of the primer pairs for the two species was demonstrated through the lack of cross-amplifications in heterologous PCR reactions and in reactions using DNA from other mycoplasma species. Subsequent restriction enzyme analysis of the amplifieduvrC gene segments from type and field strains ofM. bovisandM. agalactiaeshowed that theuvrC genes are well conserved in both species but differ significantly between the two species. The diagnostic PCR assay enabled unambiguous identification ofM. bovisandM. agalactiaestrains isolated from geographically diverse places, even in cases where 16S rRNA gene sequence analysis was unable to discriminate between the two species.