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中华实验和临床感染病杂志(电子版) ›› 2017, Vol. 11 ›› Issue (04) : 352 -358. doi: 10.3877/cma.j.issn.1674-1358.2017.04.008

临床论著

武汉地区耐碳青霉烯类肠杆菌科细菌耐药基因的研究
杨勇文1, 李从荣1,(), 李珍1, 潘月华1, 冯丽娜1, 陈浩俊1, 郭静1   
  1. 1. 430060 武汉市,武汉大学人民医院检验科
  • 收稿日期:2016-08-27 出版日期:2017-08-15
  • 通信作者: 李从荣
  • 基金资助:
    国家临床重点专科项目编号(No. 2010305)

Carbapenem-resistant Enterobacteriaceae resistant gene in Wuhan

Yongwen Yang1, Congrong Li1,(), Zhen Li1, Yuehua Pan1, Lina Feng1, Haojun Chen1, Jing Guo1   

  1. 1. Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan 430060, China
  • Received:2016-08-27 Published:2017-08-15
  • Corresponding author: Congrong Li
引用本文:

杨勇文, 李从荣, 李珍, 潘月华, 冯丽娜, 陈浩俊, 郭静. 武汉地区耐碳青霉烯类肠杆菌科细菌耐药基因的研究[J]. 中华实验和临床感染病杂志(电子版), 2017, 11(04): 352-358.

Yongwen Yang, Congrong Li, Zhen Li, Yuehua Pan, Lina Feng, Haojun Chen, Jing Guo. Carbapenem-resistant Enterobacteriaceae resistant gene in Wuhan[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2017, 11(04): 352-358.

目的

探讨武汉地区产碳青霉烯酶肠杆菌科细菌的主要耐药基因,并对耐碳青霉烯类的耐药机制进行初步分析。

方法

采用Carba NP试验对88株耐碳青霉烯类抗菌药物肠杆菌科细菌进行耐药表型确证。采用PCR方法检测KPC、NDM、VIM、IMP以及OXA-48共5种主要的碳青霉烯酶耐药基因并行测序验证。

结果

88株临床分离菌对头孢唑林和氨苄西林耐药率均高达100.0%,对哌拉西林、氨苄西林/舒巴坦、阿莫西林/克拉维酸、哌拉西林/他唑巴坦、头孢他啶、头孢噻肟、头孢吡肟和亚胺培南均> 90.0%;Carba NP实验阳性菌株76株(86.4%),其中弱阳性11株;29株检出NDM-1耐药基因,24株携带KPC-2型碳青霉烯酶耐药基因,14株检出IMP-1型耐药基因,9株检出VIM-1型耐药基因,未检出OXA-48型耐药基因;4株联合产KPC和NDM基因肺炎克雷伯菌,2株联合产NDM和VIM基因阴沟肠杆菌。

结论

本地区肠杆菌科细菌对碳青霉烯类抗菌药物的耐药机制主要是携带NDM-1和KPC-2型碳青霉烯酶基因。

Objective

To investigate the major resistance gene product carbapenemases Enterobacteriaceae in Wuhan, and to analyze the mechanism of resistance to carbapenem-resistant preliminary.

Method

Caries NP test was used to confirm the resistant phenotype of 88 strains of carbapenem-resistant antimicrobial agents Enterobacteriaceae. The KPC, NDM, VIM, IMP and OXA-48 carbapenemase resistance genes were detected by PCR and then sequenced.

Results

Total of 88 clinical isolates of cefazolin and ampicillin resistance rates were all 100.0%, while to piperacillin, ampicillin/sulbactam, amoxicillin/clavulanate, piperacillin/tazobactam, ceftazidime, cefotaxime, cefepime and imipenem were more than 90.0%. Carba NP test positive strains of 76 strains (86.4%), among which, 11 strains were weakly positive. Twenty-four strains of NDM-1 resistant gene were detected, 24 strains were resistant to KPC-2 carbapenemase, 14 strains were detected including IMP-1 resistance gene, 9 strains were detected including VIM-1 resistance gene, OXA-48 resistant gene was not detected. Four strains of Klebsiella pneumoniae combined with KPC and NDM genes, two strains of Enterobacter cloacae combined with NDM and VIM genes.

Conclusions

In Wuhan, Enterobacteriaceae carbapenem resistance mechanisms to antimicrobial agents mainly carrying NDM-1 and KPC-2 type carbapenem enzyme gene.

表1 5种主要碳青霉烯酶PCR引物信息
表2 88株分离菌对18种抗菌药物的耐药情况
图1 Carba NP实验部分结果
图2 CRE耐药基因检测部分阳性结果
图3 CRE耐药基因检测阴性对照及部分阴性结果
图4 部分OXA-48基因检测结果
图5 KPC-2测序峰图
图6 IMP-4测序峰图
图7 VIM-19测序峰图
图8 NDM-1测序峰图
表3 CRE耐药基因型(株)
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