应用多重PCR在肠杆菌科细菌中检测blaTEM、blaSHV、blaOXA1基因

Multiplex PCR detection of blaTEM, blaSHV and blaOXA1 genes in Enterobacteriaceae

摘要:

目的应用多重聚合酶链反应（PCR）法在肠杆菌科细菌中检测blaTEM、blaSHV、blaOXA1基因。方法收集2004年10月—2005年7月湖南地区中南大学3所附属医院临床标本分离的多重耐药肠杆菌科细菌171株，以美国临床实验室标准化研究所（CLSI）规定的方法进行超广谱β内酰胺酶（ESBLs）表型确证试验，多重PCR进一步进行blaTEM、blaSHV、blaOXA1三种耐药基因的检测。结果171株多重耐药的肠杆菌科细菌中，142株（83.04%）ESBLs表型检测阳性，经多重PCR扩增，105株获得阳性结果。根据扩增片段大小判断：检出携带blaTEM菌株85株，携带blaSHV菌株26株，携带blaOXA1菌株10株，其中15株菌同时携带多种耐药基因。结论多重PCR可以快速、准确地检测出ESBLs表型阳性肠杆菌科细菌是否携带blaTEM、blaSHV、blaOXA1基因。

Abstract:

ObjectiveTo apply a rapid and simple multiplex polymerase chain reaction(PCR) for detecting blaTEM，blaSHV and blaOXA-1 genes in Enterobacteriaceae. MethodsFrom October 2004 to July 2005,171 clinical strains of multiply drugresistant Enterobacteriaceae isolates were collected from three affiliated hospitals of Central South University in Hunan province. Extendedspectrum βlactamases (ESBLs)producing isolates were confirmed by means of phenotypic confirmatory tests as recommended by CLSI/NCCLS, multiplex PCR was used to determine blaTEM，blaSHV and blaOXA-1 genes. ResultsAmong 171 strains of multiply drugresistant Enterobacteriaceae,142 (83.04%) were ESBLs positive by phenotypic detection, 105 of 142 ESBLproducing Enterobacteriaceae isolates were confirmed to obtain positive results by multiplex PCR amplification. blaTEM DNA were detected in 85 isolates, blaSHV DNA in 26 isolates, blaOXA-1 DNA in 10 isolates, and 15 isolates carried  2 or more drugresistant genes.ConclusionMultiplex PCR is a suitable tool which allows the rapid genotypic detection of blaTEM, blaSHV or blaOXA1carrying bacteria from phenotypic ESBLsproducing Enterobacteriaceae.