ABSTRACT. Polymerase chain reaction (PCR) systems were developed for detection of 3 pathogenic bovine mycoplasmas, Mycoplasma alkalescens (Mak), M. bovigenitalium (Mbg) and M. bovirhinis (Mbr). The primers were selected from the sequences of the 16S rRNA from each of the mycoplasmas. Neither the Mbg-PCR system nor the Mbr-PCR system showed cross-amplification among 24 bovine, caprine-ovine and human mycoplasma species (including Acholeplasma and Ureaplasma species) tested. The Mak-PCR system showed cross-amplification with M. canadens ATCC29418 T (T=type strain). The sensitivity of each PCR system to detect the proper mycoplasma was 10 3 colony forming units (CFU) when a pure culture was tested, and 2 × 10 3 CFU when the mycoplasma culture was spiked into a calf nasal swab sample. The target mycoplasmas in a clinical nasal swab sample that could be detected by the direct plate method could also be detected by these PCR systems. -KEY WORDS: bovine mycoplasma, Mycoplasma alkalescens, M. bovigenitalium, M. bovirhinis, PCR

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