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首页> 外文期刊>Journal of Clinical Microbiology >Detection of bovine trichomoniasis with a specific DNA probe and PCR amplification system.
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Detection of bovine trichomoniasis with a specific DNA probe and PCR amplification system.

机译：用特定的DNA探针和PCR扩增系统检测牛毛滴虫病。

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Trichomoniasis is a widespread, economically important venereal disease of cattle which causes infertility and abortion. Effective control of trichomoniasis has been impeded by the insensitivity of traditional diagnostic procedures, which require the isolation and cultivation of the parasite, Tritrichomonas foetus, from infected cattle. We developed a 0.85-kb T. foetus DNA probe by identifying conserved sequences in DNAs from T. foetus that were isolated from cattle in California, Idaho, Nevada, and Costa Rica. The probe hybridized specifically to DNAs of T. foetus isolates from different geographic areas but not to DNA preparations of Trichomonas vaginalis, bovine cells, or a variety of bacteria from cattle. The probe detected DNA from a minimum of 10(5) T. foetus organisms. To improve sensitivity, a partial sequence of the probe was used to identify oligonucleotide primers (TF1 and TF2) which could be used to amplify a 162-bp product from T. foetus DNAs by PCR. A chemiluminescent internal T. foetus sequence probe was hybridized to Southern blots of the amplification product. This system detected as few as one T. foetus organism in culture media or 10 parasites in samples containing bovine preputial smegma. Analysis of 52 clinical samples showed that 47 (90.4%) of the 52 samples were correctly identified, with no false-positive reactions. In comparison, the traditional cultivation method detected 44 (84.6%) of the 52 samples from T. foetus-infected and uninfected bulls. These results indicate that the PCR-based amplification system could be a useful alternative method for the diagnosis of bovine trichomoniasis.
机译：滴虫病是一种广泛的，在经济上很重要的牛性病，可引起不育和流产。传统诊断方法的不敏感性阻碍了毛滴虫病的有效控制，传统的诊断方法需要从被感染的牛中分离和培养寄生虫，即Tritrichomonas foetus。通过鉴定来自加利福尼亚州，爱达荷州，内华达州和哥斯达黎加的牛的T.胎儿DNA的保守序列，我们开发了0.85kb T.胎儿DNA探针。该探针与来自不同地理区域的胎儿分离株的DNA特异性杂交，但不与阴道毛滴虫，牛细胞或各种牛细菌的DNA制备物杂交。该探针从至少10（5）个T.胎儿生物中检测到DNA。为了提高灵敏度，使用探针的部分序列来鉴定寡核苷酸引物（TF1和TF2），该引物可用于通过PCR从胎儿T. DNA扩增162 bp的产物。将化学发光的内部T.胎儿序列探针与扩增产物的Southern印迹杂交。该系统在培养基中检测到少至一种T.胎儿生物，在含有牛原皮包皮垢的样品中检测到10种寄生虫。对52个临床样本的分析表明，在52个样本中有47个（90.4％）被正确识别，没有假阳性反应。相比之下，传统的耕种方法在感染了费城犬的公牛和未感染公牛的52个样本中检测到了44个（84.6％）。这些结果表明，基于PCR的扩增系统可能是诊断牛滴虫病的一种有用的替代方法。

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