Differentiation of Mycoplasma bovis, Mycoplasma bovigenitalium, Mycoplasma californicum and identification of Ureaplasma diversum by real-time PCR

Mycoplasmas and ureaplasmas are important etiological agents of mastitis, pneumonia and reproductive disorders in cattle, which cause significant economic damage to cattle farming. The most significant species are Mycoplasma bovis , M. bovigenitalium , M. californicum and Ureaplasma diversum . Commercial diagnostic PCR systems for the detection of bacteria of the genus Mycoplasma in different biological samples are described, but no PCR kits have been developed to address the identification of Mycoplasma species. In this work, real time PCR assays for differentiation of pathogenic mycoplasmas ( Mycoplasma bovis, M. bovigenitalium, M. californicum ) and detection of Urea-plasma diversum in biological material (semen, milk, vaginal swabs, tissues) are developed. UvrC gene for M . bovis , 16S rRNA gene for M. bovigenitalium and U. diversum , and rpoB gene for M. californicum were chosen as target genes. The PCR assays included a system of primers and probes for detection of exogenous noncompetitive internal control sample...