牛环形泰勒虫病二温式PCR诊断方法的建立及初步应用  

Establishment and Primary Application of A Two-temperature PCR Method for DetectionTheileria annulata Infection in Cattle

作　　者：王振宝[1,2] 刘启生[2] 哈森 何晓杰[3] 刘绪斌 孟茹 叶尔保勒 巴音查汗[2] 

机构地区：[1]伊犁出入境检验检疫局综合技术服务中心,新疆伊宁835000 [2]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [3]伊犁职业技术学院,新疆伊宁835000

出　　处：《动物医学进展》2012年第5期59-63,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(30960282)

摘　　要：为建立特异、敏感、快速的二温式PCR诊断方法,根据牛环形泰勒虫裂殖子表面抗原(tams1)基因,设计了一对特异性引物,扩增出大小为154bp基因片段,经克隆、测序分析,与已知基因序列的相似性为96%。用建立的牛环形泰勒虫病二温式PCR诊断方法,对从新疆牛环形泰勒虫病流行地区采集的50份全血样品进行诊断,阳性率为88%,而血涂片检出的阳性率只有58%。经试验验证,该方法具有特异性高、敏感性强、重复性和稳定性好等优点。表明本试验所建立的二温式PCR诊断方法可用于牛环形泰勒虫病的临床诊断、隐性感染检测和流行病学调查。To establish a specific,sensitive and rapid two-temperature PCR assay for detection of Theileria annulatainfection in cattle.The primers were designed according to T.annulata merozoite surface antigen（Tams1）,and a gene fragment of 154bp was amplified by the two-temperature PCR.The cloning and sequencing results showed that the sequence of PCR products had 96%similarity to the Tams1.Fifty blood samples collected from cattle in Theileria annulata epidemic area of Xinjiang were detected.Out of the 88%samples were positive by two-temperature PCR assay,while the positive rate was only 58% detected from same carrier cattle by blood smear examination.The results indicated that this assay could be widely used to clinical diagnosis or suppressive infection and epidemiological investigation for Theileria annulata infection in cattle.

关 键 词：牛 环形泰勒虫 二温式PCR 克隆 诊断 应用 

分 类 号：S855.91[农业科学—临床兽医学]