牛卵形巴贝斯虫病二温式PCR诊断方法的建立  

Establishment of Two-temperature PCR Method for Detection Babesia ovata in Bovine

作　　者：田万年 王妍慧 薛书江[2] 贾立军[2] 张守发[2] TIAN Wan-nian;WANG Yan-hui;XUE Shu-jiang;JIA Li-jun;ZHANG Shou-fa(College of Animal Science,Jilin Agricultural Science and Technology College,Jilin 132101,China;College of Agriculture,Yanbian University,Yanji 133002,China)

机构地区：[1]吉林农业科技学院动物科技学院,吉林吉林132101 [2]延边大学农学院,吉林延吉133002

出　　处：《中国兽医杂志》2020年第9期38-40,共3页Chinese Journal of Veterinary Medicine

基　　金：吉林农业科技学院大学生科技创新项目(2018114-39026号);吉林省教育厅“十三五”科学技术项目(JJKH20190981KJ)。

摘　　要：为建立一种特异、敏感、快速的牛卵形巴贝斯虫病诊断方法。根据GenBank已报道的牛卵形巴贝斯虫18S rRNA基因(LC125457)设计合成了1对引物,通过条件优化,建立了牛卵形巴贝斯虫病二温式PCR诊断方法。该方法能扩增出464 bp的牛卵形巴贝斯虫特异性基因片段,测序结果与已知基因序列同源性为100%。对牛卵形巴贝斯虫基因组DNA的最小检测量为16 fg/μL,与牛瑟氏泰勒虫、牛巴贝斯虫和双芽巴贝斯虫均无交叉反应。二温式PCR诊断方法具有较高的特异性和敏感性,可用于牛卵形巴贝斯虫病的快速诊断和流行病学调查。To establish a rapid,specific and sensitive method for detection of Babesia ovata in bovine,a pair of specific primers were designed and synthesized according to 18S rRNA gene in GenBank(LC125457)and a two-temperature PCR assay was established.A 464 bp gene fragment was amplified,which showed 100%of homology with the known gene sequence.The minimum detectable amount of DNA was 16 fg/μL.The method showed no cross reaction with Theileria sergenti,Babesia bovis,and Babesia bigemina.The results showed that the PCR method was specific and sensitive.This study provides a method for rapid clinical diagnosis and epidemiological investigation of Babesia ovata in bovine.

关 键 词：卵形巴贝斯虫 18S rRNA基因 二温式PCR 

分 类 号：S852.723[农业科学—基础兽医学]