【作者】 程文娟； 彭少华；

【Author】 Shao-hua Peng,(Department of laboratory science,Renmin Hospital of Wuhan University,Wuhan 430060,China)

【机构】 武汉大学人民医院检验科；

【摘要】 目的探讨多重PCR(multiplex polymerase chain reaction)方法用于同时检测耐甲氧西林金黄色葡萄球菌(Methicillin resistant Staphylococcus aureus,MRSA)及其对氨基糖苷类抗生素耐药性的可行性。方法优化多重PCR条件,同时检测金黄色葡萄球菌(SA)的三个基因。应用此方法检测212株临床分离菌株,用盲法试验检测14株未知菌,并进行统计学分析和评价。结果多重PCR受多种因素影响,其产物经电泳、测序、分析为目的片断。以传统方法鉴定结果为金标准,Kappa检验表明多重PCR鉴定结果与金标准无显著性差异,一致性好,阳性似然比和阴性似然比均好。讨论多重PCR可用于检测MSSA、MRSA、对氨基糖苷类抗生素耐药的MRSA,不宜用于检测对氨基糖苷类抗生素耐药的MSSA。更多还原

【Abstract】 Objective We explored to developed a multiplex PCR(mPCR)for the purpose of identifying Staphylococcus aureus,mecA gene(encoding Methicillin resistant)and aac(6’)/aph(2")enzyme(gentamicin resistant).Methods A mPCR assay was developed to simultaneously detect three genes of Staphylococcus aureus.We identified 212 clinical isolates by the mPCR,and subsequently evaluated for its effectiveness.Results The result showed that we get the expected target DNA framents and the mPCR assay could attribute to multiple factors.We regarded the routin assay as"gold standard",the results of Kappa test indicated that the result of mPCR assay were the same as the routin assay and the likelihood ratios showed the mPCR could be used as diagnosis test.Conclusions A simple,rapid,multiplex PCR technique has now been developed for the simultaneous detection nuc gene、mecA gene and aac(6’)/aph(2")gene of Staphylococcus aureus could be helpful for clinical diagnosis.更多还原