痰标本中金黄色葡萄球菌的分离鉴定方法研究

Identify apathogenic staphylococcus aureus from a sputum specimen

目的探索一种基于16SrRNA基因的细菌快速鉴定方法,为临床标本中未知病原菌的诊断提供新的方法思路。方法对1份临床发热伴呼吸道症候群患者的痰标本进行分离培养,获得3种不同的菌落。直接以纯菌落为模板,以通用引物扩增未知菌的16SrRNA基因片段,产物直接测序后进行基于局部比对算法的搜索工具比对,根据序列同源性鉴定病原细菌。结果 3种菌落分别为不易致病的表皮葡萄球菌、龋齿罗特放线菌及致病性的金黄色葡萄球菌,后者继续用特异性引物扩增检测鉴定,确认为金黄色葡萄球菌。结论本研究建立了一种利用16SrRNA基因扩增快速鉴定临床标本中未知病原菌的简便方法。

Objective To explore a rapid bacteria identified method based on the analysis technology of 16S rRNA gene sequence for the diagnosis of the clinical specimens .Methods Three kinds of pure colonies were isolated and cultured from the sputum specimen sample from a patient with fever and respiratory syndrome .The colonies were used as templates ,universal primers were used to amplify 16S rRNA gene fragments of unknown bacteria ,then se-quenced the products directly ,and compared using the Basic Local Alignment Search Tool （ BLAST ） .Results A strain of Staphylococcus epidermidis ,a strain of Rothia dentocariosa and a strain of Staphylococcus aureus were iden-tified ,and the last was confirmed by specific primers .Conclusion This study developed a simple method for rapid i-dentification of pathogens from clinical specimens by using 16S rRNA gene amplification .