牛新孢子虫病PCR检测方法的建立和应用

Development and application of polymerase chain reaction assay for detection of neospora disease in bovine

犬新孢子虫感染是导致妊娠母牛流产的主要原因之一,准确快速地诊断是有目的治疗该病的前提。本研究根据已知的犬新孢子虫种属特异性基因片段Nc-5基因序列,设计了一对特异性引物,建立了检测犬新孢子虫的PCR技术。利用本方法可以从感染牛胎儿的脑脊液及实质脏器中扩增出1条大小为350 bp的特异性核酸片段。在吉林省延边龙井地区的应用检测结果表明,流产牛胎儿中新孢子虫感染率为17%(15/90)。本方法在实际应用中快速、灵敏、特异性高,可以用于牛和其它动物新孢子虫病的快速诊断和流行病学调查。

Neospora can/hum infection is one of the main cause of miscarriage in pregnant cows. Successful treatment relies on early and fast diagnosis. Double differential primers were designed according to neospora eaninum NC-5 gene sequence of the United States strain and PCR was established for the detection of neospora caninum. A 350 bp differential nuclear acid fragment was amplified from eerebrospinal fluid and organs of the infection earle embryo. Using this method, 17 % （15/90） earle embryo of miscarriages was detected neospora eaninum positive in the Longjing region of Yanbian. This method is fast and has high sensitivity ＆ specificity and could be useful in epidemiology study of neospora eaninum infection in earle and other animals.