【摘要】 目的研究聚合酶链反应(PCR)和高分辨熔解技术(HRM)两种方法检测细菌耐药基因氨基糖苷乙酰转移酶的变异基因(aac(6′)-Ib-cr)检出率的差异性。方法采用PCR方法检测299株临床常见的革兰阴性杆菌的aac(6′)-Ib基因,并以内切酶BtsCI酶切消化aac(6′)-Ib的PCR阳性产物以确定aac(6′)-Ib-cr;同时采用HRM技术检测以上所有菌株的aac(6′)-Ib和aac(6′)-Ib-cr基因。结果 PCR和酶切法共检出aac(6′)-Ib基因29株,其中有21株变异为aac(6′)-Ib-cr,变异率为72.4%(21/29)。而且HRM技术检测的结果与其完全一致。结论 PCR和HRM两种方法检测细菌耐药基因aac(6′)-Ib-cr的能力相当,可以用更简便的HRM技术替代PCR和酶切法检测aac(6′)-Ib-cr基因,尤其适用于大量筛检。更多还原

【Abstract】 Objective To study the difference of detection rate between PCR and high-resolution melt(HRM)for detecting the bacterial drug resistance gene aac(6′)-Ib-cr.Methods The PCR method was adopted to detect the aac(6′)-Ib gene in 299 strains of common Gram-negative bacilli,and the PCR positive products of aac(6′)-Ib were digested with BtsCI.Meanwhile Aac(6′)-Ib and aac(6′)-Ib-cr genes in above bacterial strains were detected by adopting the HRM technique.Results A total of 29 isolates were identified as aac(6′)-Ib gene,and 21 mutations were aac(6′)-Ib-cr,with a mutation rate of 72.4%(21/29)for PCR and restriction enzyme digestion.And the results of the HRM test were in good agreement with each other.Conclusion PCR and HRM are similar for the detection of the bacterial resistance gene aac(6′)-Ib-cr,and aac(6′)-Ib-cr gene could be detected by a simpler HRM technique instead of PCR and enzyme digestion,especially for a large number of screening.更多还原