多重PCR检测肠杆菌科细菌碳青霉烯类耐药基因型  

Application of multiplex PCR in detection of genotypes of carbapenem-resistant Enterobacteriaceae

作　　者：刘颖[1] 张会英[1] 郭宇[1] 王艳[1] 梁倩 童静[1] 吴俊[1] LIU Ying;ZHANG Hui-ying;GUO Yu;WANG Yan;LIANG Qian;TONG Jing;WU Jun(Beijing Jishuitan Hospital,Beijing 100035,China)

机构地区：[1]北京积水潭医院检验科,北京100035

出　　处：《中华医院感染学杂志》2020年第9期1281-1285,共5页Chinese Journal of Nosocomiology

基　　金：吴阶平医学基金会临床科研专项基金资助项目(320.6750.10078)。

摘　　要：目的采用多重PCR技术快速、高效检测耐碳青霉烯类肠杆菌科细菌(CRE)的耐药基因型,为CRE的预防和控制提供依据。方法收集北京积水潭医院住院患者分离得到的58株CRE菌株。CRE菌株表型检测采用改良Hodge试验和亚胺培南/亚胺培南联合EDTA E-test协同试验进行。CRE菌株基因型检测采用多重PCR扩增反应体系(包括11对碳青霉烯酶基因引物:A组为blaIMP,blaSPM,blaVIM,B组为blaBIC,blaNDM,blaKPC,blaOXA,C组为blaAIM,blaGIM,blaSIM,blaDIM)。对blaKPC阳性的肺炎克雷伯菌行全酶基因编码区PCR扩增及产物测序。结果58株CRE菌株中52株为肺炎克雷伯菌,占89.66%;3株为大肠埃希菌,占5.17%;2株为阴沟肠杆菌,占3.45%;1株为弗劳地枸橼酸杆菌,占1.72%。改良Hodge试验阳性52株(89.66%),亚胺培南/亚胺培南联合EDTA E-test协同试验阳性2株(3.45%)。CRE中检出blaIMP基因型1株(1.72%),为肺炎克雷伯菌;blaKPC基因52株(89.66%),其中肺炎克雷伯菌为50株(86.21%),2株为大肠埃希菌(3.45%);blaNDM基因1株(1.72%),为阴沟肠杆菌;11种基因检测全阴性为4株(6.90%),分别为肺炎克雷伯菌、阴沟肠杆菌、大肠埃希菌、弗劳地枸橼酸杆菌各1株,基因检测阳性者均无重叠。blaKPC基因阳性的肺炎克雷伯菌全酶基因编码区测序结果为blaKPC-2亚型。结论北京积水潭医院患者检出的CRE中,以肺炎克雷伯菌为主。共检测出3种基因型,其中blaKPC-2型是医院CRE的主要基因型,提示医院感控部门要加强对肺炎克雷伯菌耐药的监控,避免耐药率进一步上升。OBJECTIVE To employ a multiple PCR technology for rapid and efficient detection of genotypes of carbapenem-resistant Enterobacteriaceae(CRE)strains isolated from Beijing Jishuitan Hospital so as to provide guidance for prevention and control of CRE.METHODS Totally 58 strains of CRE are isolated from specimens of hospitalized patients.The phenotypes of CRE strains were confirmed by modified Hodge test and imipenem/imipenem combined with EDTA E-test synergy test,the genotypes of the CRE strains were detected by 3 group of multiplex PCR reactions,including 11 pairs of primers:group A-blaIMP,blaSPM,blaVIM;group B-blaBIC,blaNDM,blaKPC,blaOXA;group C-blaAIM,blaGIM,blaSIM,blaDIM.PCR reaction and DNA sequencing of complete KPC-coding zone were performed for blaKPC-positive Klebsiella pneumoniae strains.RESULTS Among the 58 strains of CRE,there were 52 strains of K.pneumoniae(89.66%),3 strains of Escherichia coli(5.17%),2 strains of Enterobacter cloacae(3.45%),1 strain of Citrobacter freundii(1.72%).Totally 52(89.66%)strains showed positive for modified Hodge test,and 2(3.45%)strains were positive for imipenem/imipenem combined with EDTA E-test synergy test.Among the isolated CRE strains,there were 1 strain of K.pneumoniae carrying with blaIMP,52(89.66%)strains carrying with blaKPC,including 50(86.21%)strains of K.pneumoniae and 2(3.45%)strains of E.coli,and 1(1.72%)strain of E.cloacae carrying with blaNDM.Totally 4(6.90%)strains were tested negative for 11 genotypes,including 1 strain of K.pneumoniae,1 strain of E.cloacae,1 strain of E.coli and 1 strain of Citrobacter freundii;there was no overlap in genotype-positive results.The blaKPC-positive K.pneumoniae strains were confirmed as the blaKPC-2 subtype by DNA sequencing of complete KPC-coding zone.CONCLUSION K.pneumoniae strains are dominant among the CRE strains isolated from the patients of Beijing Jishuitan Hospital.Totally 3 genotypes have been detected,blaKPC-2 is the major genotype of the CRE strains,indicating that it is necessary for the infection control depar

关 键 词：肠杆菌细菌 碳青霉烯酶 基因型 

分 类 号：R446.5[医药卫生—诊断学]