通用引物PCR-SSCP技术分析16SrRNA基因特征快速鉴定细菌

Analysis of 16SrRNA gene by Universal primer polymerase chain reaction single - strand conformation polymorphism to identify bacteria rapidly.

目的探讨通用引物聚合酶链反应-单链构象多态性(PCR-SSCP)技术在细菌检测中的应用。方法选取9种常见的细菌,采用通用引物对细菌的16SrRNA基因进行PCR扩增,产物进行单链构象多态性分析,并以标准菌株为对照。结果一对引物扩增产物SSCP图谱各细菌之间难以区别;两对引物扩增产物SSCP图谱条带数目、相对迁移率及条带之间的间距存在明显,可相互区别;3种细菌的标准菌株和临床菌株的SSCP图谱完全一致。结论采用针对16SrRNA的通用引物PCR-SSCP技术以标准菌株为对照可方便快捷地检测细菌。

Objective To discuss use of universal primer polymerase chain reaction single-strand conformation polymorphism (PCR-SS-CP) analysis in identification of bacteria.Methods The 16S ribosomal RNA genes of 9 frequent bacteria were amplified by universal primer polymerase chain reaction,men PCR products were analyzed by single-strand conformation polymorphism in comparison with standard bacteri-a.Results The SSCP patterns of PCR products which were amplified by one primer set were difficult to distinguish,but numbers relative and transfer rate and space of strips in the SSCP patterns which were amplified by two pnmer sets were clearly different and could be identified.The SSCP patterns of standard bacteria accorded with clinical bacteria.Conclusion Universal primer PCR aimed at 16SrRNA can rapidly and conveniently identify bacteria in comparison with standard bacteria.