期刊论文

中文

中国兽医科学

1673-4696

2010

40

12

1255-1258

10.16656/j.issn.1673-4696.2010.12.010

湖南省科技计划项目（2010NK3015）；

本校为第一机构

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 th...