Kobayashi H, Hirose K, Worarach A, Paugtes P, Ito N, Morozumi T, Yamamoto K
National Institute of Animal Health, Ibaraki, Japan.
J Vet Med Sci. 1998 Dec;60(12):1299-303. doi: 10.1292/jvms.60.1299.
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 ATCC29418T (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 x 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.
已开发出聚合酶链反应（PCR）系统用于检测3种致病性牛支原体，即碱性支原体（Mak）、牛生殖支原体（Mbg）和牛鼻支原体（Mbr）。引物是从每种支原体的16S rRNA序列中选取的。在所测试的24种牛、山羊-绵羊和人支原体物种（包括无胆甾原体和脲原体物种）中，Mbg-PCR系统和Mbr-PCR系统均未出现交叉扩增。Mak-PCR系统与加拿大支原体ATCC29418T（T = 模式菌株）出现交叉扩增。当检测纯培养物时，每个PCR系统检测相应支原体的灵敏度为10³菌落形成单位（CFU），当将支原体培养物加入小牛鼻拭子样本中时，灵敏度为2×10³CFU。通过直接平板法可在临床鼻拭子样本中检测到的目标支原体，也可通过这些PCR系统检测到。
