粪肠球菌PCR检测方法的建立  

Establishment of a PCR assay for detection of Enterococcus faecalis

作　　者：黎满香[1] 林荣高[1] 薛立群[1] 蒋伟[1] 陈滔[1] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128

出　　处：《中国兽医科学》2010年第12期1255-1258,共4页Chinese Veterinary Science

基　　金：湖南省科技计划项目(2010NK3015)

摘　　要：为建立一种基于细菌16 S rDNA的粪肠球菌PCR检测方法,用于检测临床样本和鉴定分离纯化的细菌,通过比较肠球菌间16 S rDNA的差异,设计了1对特异性引物,并对PCR反应条件进行了优化,对PCR的特异性和敏感性进行了检测。结果显示,粪肠球菌ATCC29212及42个鉴定阳性的临床分离株为PCR阳性,检测结果与16 S rDNA测序的鉴定结果一致,其他种属的4株菌为PCR阴性,所得扩增条带单一且与预期的产物大小一致,测序表明该条带为目的条带;该方法的最小检出量为134 CFU,并能直接对病料进行检测。表明所建立的PCR方法能特异地检测粪肠球菌,且敏感、简易、快捷,适用于实验室的日常检测。The purpose of the present study was to develop a PCR assay based on bacterial 16S rDNA for the routine detection of Enterococcus faecalis.A pair of specific primers was designed by comparing the 16S rDNA of Enterococcus species.Optimization of PCR condition,specificity and sensitivity were conducted to evaluate the PCR assay.The results showed that the PCR result of ATCC29212 and 42 clinical isolates determined previously by 16S rDNA sequencing was positive,which accorded with the 16S rDNA sequencing,but other genus was negative.A single segment was amplified for those positive results and the sequencing result confirmed that the segment was the target fragment.In addition,the PCR assay could detect clinical samples directly,requiring no bacterial culture and the detection limit was 134CFU.The method was sensitive,accurate and rapid,and was suitable for the routine laboratory detection of E.faecalis.

关 键 词：粪肠球菌 聚合酶链反应 检测 

分 类 号：S852.611[农业科学—基础兽医学]