驽巴贝斯虫二温式PCR检测方法的建立及初步应用  

Establishment and Application of Two-temperature PCR for Babesia caballi Detection

作　　者：张杨[1] 达伊力特 刘进 李佳[1] 巴音查汗[1] 

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [2]新疆和静县巴音布鲁克镇兽医站,新疆和静841300 [3]新疆和静县畜牧兽医站,新疆和静841300

出　　处：《动物医学进展》2014年第6期16-20,共5页Progress In Veterinary Medicine

基　　金：国家科技支撑计划项目(2012BAD46B01-04)

摘　　要：为了建立一种能快速、特异、敏感地检测马驽巴贝斯虫的方法,根据GenBank中的驽巴贝斯虫(Babesia caballi)Bc-48基因序列设计了一对特异性引物,建立检测驽巴贝斯虫的二温式PCR方法。该方法能够特异地扩增155bp的片段,与双芽巴贝斯虫、环形泰勒虫、马泰勒虫、瑟氏泰勒虫均无交叉反应,能检测到的最低DNA拷贝数为5.17×102 copies/μL。应用所建立的方法检测了采集于和静县焉耆马匹样品,结果显示,马驽巴贝斯虫感染率为69.51%(57/82),与血液涂片检查结果37.8%(31/82)相比,其检出率更高。表明该二温式PCR检测方法具有较高的特异性和敏感性,可用于马驽巴贝斯虫病的早期诊断。To establish a rapid,specific and sensitive two-temperature PCR assay for detection of Babesia caballi ,specific primers were designed according to Bc-48 gene of Babesia caballi published in GenBank.A gene fragment of 155 bp was amplified by this method,and there was no cross reaction with B.bigemina, T.annulata,T.equi,T.sergenti,and the lowest detection limit of Babesia caballi DNA was 5.17 × 10 ^2 copies/μL.Eighty-two blood samples collected from horses in Hejing were detected by the established method.The results showed that the positive rate was 69.51%（57/82）,while that of blood smear test was 37.8%（31/82）.This study indicated that established two-temperature PCR method could be used for early diagnosis of B.caballi infection with its high specificity and sensitivity.

关 键 词：马驽巴贝斯虫 Bc-48 基因 样品检测 

分 类 号：S852.723[农业科学—基础兽医学]