牛瑟氏泰勒虫ITS基因PCR诊断方法的建立  

Establishment of PCR Method for the Detection of Theileria sergenti Based on ITS Gene

作　　者：王轶男[1] 贾立军[1] 薛书江[1] 张影[1] 钱年超[1] 张守发[1] 

机构地区：[1]延边大学农学院动物医学系,吉林延吉133002

出　　处：《中国畜牧兽医》2012年第8期80-82,共3页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(30960278)

摘　　要：本试验根据GenBank上登录的牛瑟氏泰勒虫ITS基因序列(AY661522.1),应用Primer Premier 5.0和Oligo 6.31软件设计合成1对特异性引物,以牛瑟氏泰勒虫DNA为模板,建立了牛瑟氏泰勒虫ITS基因PCR诊断方法。该方法扩增片段大小为1020 bp,与参考序列的同源性为98%;建立的PCR方法与猪附红细胞体、犬新孢子虫和弓形虫均无交叉反应,最低DNA检出量为1.5 pg/μL;通过对60份临床样品的检测,并与血涂片方法进行比较,结果显示PCR方法具有特异、敏感等特点,适用于牛瑟氏泰勒虫的检测。According to ITS gene sequence of Theileria sergenti in GenBank（AY661522.1）,a pair of specific primers was designed and synthesized using Primer Premier 5.0 and Oligo 6.31.Using T.sergenti DNA as templates,PCR method of T.sergenti was established.1020 bp fragments were amplified.The fragments shared 98% homology with the reference sequence.The established PCR method showed no cross reaction with Mycoplasma suis,Neospora caninum and Toxoplasma gondii RH strain.The minimum detectable DNA amount was 1.5 pg/μL.Sixty clinical samples were detected by PCR method and microscopic examination of Giemsa-stained blood smears.The results showed that the PCR method was sensitive and specific.It was suitable for the detection of T.sergenti.

关 键 词：牛瑟氏泰勒虫 ITS基因 PCR 

分 类 号：S854.43[农业科学—临床兽医学]