检验医学与临床2018，Vol.15Issue(9)：1235-1238,4.DOI:10.3969/j.issn.1672-9455.2018.09.006
耐碳青霉烯肺炎克雷伯菌的耐药机制研究
Mechanisms of carbapenem-resistant Klebsiella pneumoniae
摘要
Abstract
Objective To investigate the molecular mechanism of carbapenem-resistant Klebsiella pneu-moniae(CRKP)isolated from our hospital in 2014.Methods Imipenem non-sensitive strains were screened by disk diffusion method or Vitek 2 Compact automatic susceptibility analysis system and confirmed by agar dilu-tion method.Carbapenemases phenotypes was confirmed by Carba NP test.Carbapenem resistance genes,su-perwide-spectrum β lactamase(ESBLs)genes and plasmid-carrying AmpC gene were amplified by PCR and sequenced.The changing of outer membrane protein of the strain was analysised by SDS-PAGE methods.Ef-flux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone(CCCP)inhibition test was used to detected the role of efflux pump in carbapenem-resistance Klebsiella pneumoniae.Results A total of strains of carbap-enem-resistant strains were confirmed by agar dilution method,and 19 strains producing KPC-2 carbapenemase strain of which were Carba NP phenotype positive 18 stranis producting KPC-2,1 stain producting NOM-1. The rate of blaCTX-M,blaSHV,blaDHA blaTEM in the strains which producing no carbapenemase were 72.7%,27.3%,27.3% and 72.7%.The results of SDS-PAGE showed that only the OmpK36 protein of strain No.2368,No.2875,No.3050 were lost.In the presence of CCCP,the minimum inhibitory concentration(MIC) of Imipenem,Meropenem and Ertapenem in the 11 KPC2-producing strans was significantly descend.Conclu-sion The production of carbapenemase,especially KPC-2 is CRKP to carbapenem antibiotics in Klebsiella pneumoniae,followed by high expression of efflux pump and the production of AmpC.关键词
肺炎克雷伯菌/碳青霉烯酶基因/SDS-PAGE/羰酰氰氯苯腙抑制试验Key words
Klebsiella pneumoniae/carbapenemase gene/SDS-PAGE/efflux pump carbonyl cyanide 3-chlorophenylhydrazone inhibition test分类
医药卫生引用本文复制引用
程莉,谭婷婷,魏红霞,张葵..耐碳青霉烯肺炎克雷伯菌的耐药机制研究[J].检验医学与临床,2018,15(9):1235-1238,4.基金项目
江苏省南京市医学科技发展资金资助项目(ZKX13028). （ZKX13028）
