驽巴贝斯虫病PCR检测方法的建立和应用  

Application and Development of PCR Assay to Detect Babesia caballi

作　　者：罗金[1,2] 刘光远[2] 谢俊仁[2] 田占成[2] 沈辉[2] 王芳芳[2] 孙晓林[1] 陈荣贵 王海军 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室农业部草食动物疫病重点开放实验室,甘肃兰州730046 [3]新疆伊犁州动物疾病控制与诊断中心,新疆伊犁735000 [4]新疆新源县畜牧兽医站,新疆新源830000

出　　处：《中国畜牧兽医》2011年第7期193-195,共3页China Animal Husbandry & Veterinary Medicine

基　　金：甘肃省科技重大专项(092NKDA031)

摘　　要：根据驽巴贝斯虫(Babesia caballi)18S rRNA基因序列设计1对特异性引物,扩增出452 bp核苷酸片段,建立了检测驽巴贝斯虫病的PCR方法。敏感性试验结果表明,该方法最低能检出0.01 fg/μL驽巴贝斯虫DNA模板。特异性试验结果显示,在被检测的6个巴贝斯虫株中,仅驽巴贝斯虫株能扩增出特异性片段,马泰勒虫、双芽巴贝斯虫、莫氏巴贝斯虫、卵形巴贝斯虫、大巴贝斯虫的扩增结果均为阴性。对45份马属动物血样进行检测,本研究建立的PCR方法测得驽巴贝斯虫病的阳性率为26.67%(12/45),与显微镜检测方法进行了比较,结果显示PCR检测方法可显著提高驽巴贝斯虫的检出率。The 18S rRNA gene recently discovered was shown to be species-specific.A pair of primers was designed to specifically amplified a 452 bp fragment.The PCR result of specificity assay showed that one references B.caballi could be detected by the PCR test,but no amplification was observed when other five bacterial species isolated from T.equi,B.bigimina,B.motasi,B.ovata,B.major tissue were detected.And the sensitivity result showed that the minimum dose of B.caballi that could be detected by PCR assay was 0.01 fg/μL,45 clinical sample,from horses farm in China,doubtedly infected with B.caballi were tested by PCR.The results showed that 12 positive samples could be detected by the PCR assay.At the same time,microscope were also used for the clinical samples.The test revealed that the sensitivity of the PCR assay was higher than that the microscope test.

关 键 词：驽巴贝斯虫 PCR诊断方法 应用 

分 类 号：S852.72[农业科学—基础兽医学]