牛瑟氏泰勒虫病PCR诊断方法的建立  

Establishment of a PCR Assay on Theleria sergenti in Cattle

作　　者：白光彦 杨艳玲[1,2] 张西臣[2] 尹继刚[3] 陈峙峰[2] 

机构地区：[1]吉林市丰满区小白山乡畜禽防疫服务中心,吉林丰满132108 [2]吉林大学畜牧兽医学院,长春130062 [3]吉林大学人兽共患病研究所,长春130062

出　　处：《寄生虫与医学昆虫学报》2011年第1期48-51,共4页Acta Parasitologica et Medica Entomologica Sinica

摘　　要：为建立一种快速、敏感的牛瑟氏泰勒虫病诊断方法,本实验分离提取牛瑟氏泰勒虫全血基因组DNA,并设计一对编码牛瑟氏泰勒虫32 kDa （P32） 主要表面蛋白基因的引物,进行PCR扩增,预期扩增的片段为875 bp.并对该实验进行特异性、敏感性及临床检测实验.结果成功扩增出了长为875 bp的基因片段,且具有高特异性和很好的敏感性,表明该方法可用于牛瑟氏泰勒虫感染的临床检测.A PCR test was developed to detect Theleria sergenti in cattle which proved to be a quick and sensitive methods. A pair of synthetic oligonucleotide primers, designed from the gene encoding a 32 kDa surface protein of T. sergenti, was used to amplify parasite DNA from the blood of T. sergenti-infected cattle by the Polymerase Chain Reaction （PCR）. The results showed that a predicted 875 bp DNA product was achieved. The detection with PCR for T. sergenti is specific because of no amplification in B. equi, bovine leukocyte and T. annulata. As shown in results of sensitivity for PCR, the minimal number of DNA copies was 4.5 according to the maximal template DNA dilution of 1 ： 100 000. Moreover, of 50 specimens from grazing cattle, 18 were microscopically positive, whereas PCR revealed 31 samples were positive. Therefore, PCR provides a useful diagnostic tool for detecting T. sergenti infected cattle, and it is significantly more sensitive than the current microscope methods.

关 键 词：牛瑟氏泰勒虫 PCR 基因组DNA P32 诊断方法 

分 类 号：Q78[生物学—分子生物学]