论文部分内容阅读
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。
Objective To investigate the difference of the detection rates of aac (6 ’) - Ib-cr between the resistant gene aminoglycoside acetyltransferase and polymerase chain reaction (PCR) and high resolution melting (HRM) . Methods The aac (6 ’) - Ib gene of 299 clinically common Gram - negative bacilli was detected by PCR and the PCR positive products of aac (6’) - Ib were digested with restriction endonucleases BtsCI to determine aac (6 ’ -Ib-cr; aac (6 ’) - Ib and aac (6’) - Ib-cr genes of all the above strains were also detected by HRM technique. Results 29 strains of aac (6 ’) - Ib gene were detected by PCR and digestion. Among them, 21 strains were aac (6’) - Ib-cr with mutation rate of 72.4% (21/29). And HRM technology test results and exactly the same. Conclusion The PCR and HRM methods are comparable in detecting the resistance gene aac (6 ’) - Ib-cr. The simpler HRM technique can be used to detect aac (6’) - Ib-cr gene instead of PCR and restriction enzyme digestion. Especially suitable for mass screening.