中文摘要：

目的建立一种适于临床微生物实验室快速检测blaKPC与blaNDM基因的多重PCR方法。方法根据Gene Bank提供的序列,人工合成blaKPC与blaNDM基因模板以及多重PCR反应的引物;利用产KPC的大肠埃希菌作为阳性对照,对新构建的多重PCR反应进行验证,并且对临床15例临床分离鉴定的疑似产KPC与NDM的耐药细菌进行检测。结果成功构建了用于检测blaKPC与blaNDM基因的多种PCR反应体系,可以筛查出产KPC的大肠埃希菌;检测的15株临床分离鉴定的细菌均为阴性。结论构建的多重PCR反应体系可以快速准确地对临床产KPC与NDM细菌进行检测,以用于临床疑似细菌的进一步确认。

英文摘要：

OBJECTIVE To develop a rapid and reliable multiplex PCR-based technique for detection of bla KPC and blaNDM genes in clinical laboratory. METHODS Templates of blaKPC or blaNDM and primers to amplify blaKPC or blaNDM were designed according to Gene Bank. And DNA extracted from KPC-producing Escherichia coli was used as positive to confirm the specificity of multiplex PCR, totally 15 cases of clinically isolated drug resistant bacteria which were suspected to produce KPC and NDM were detected. RESULTS We have successfully developed a rapid and reliable multiplex PCR system for the detection of bzoKPC or blaNDM, which can screen out the KPC- producing E. coli; all the 15 clinical isolates were negative. CONCLUSION. Multiplex PCR reaction system can detect the KPC or NDM-producing bacteria, which therefore can be applied for the further identification of the clinical suspected bacteria.