儿童患者中分离的碳青霉烯类耐药肺炎克雷伯菌分子流行病学分析及耐药机制研究

发布时间：2018-07-06 10:14

本文选题：儿童 + 碳青霉烯酶　；参考：《中国感染与化疗杂志》2016年05期

【摘要】：目的研究儿童患者中分离的碳青霉烯类耐药肺炎克雷伯菌(CRKP)的流行特征及耐药机制。方法收集2013年1-12月上海市儿童医院临床微生物室分离的12株碳青霉烯类耐药肺炎克雷伯菌,采用琼脂稀释法检测其对常用抗菌药物的耐药性,乙二胺四乙酸(EDTA)协同试验和3'-氨基苯硼酸(APB)协同试验进行碳青霉烯酶表型分析,PCR扩增及DNA测序进行碳青霉烯酶基因型确证,接合试验检测碳青霉烯酶耐药基因是否位于质粒上,脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)检测菌株间基因相关性。结果 12株CRKP对多黏菌素E均敏感,对头孢菌素类耐药率均为100%,对亚胺培南、美罗培南和厄他培南耐药率分别为91.7%、91.7%和100%。PCR及DNA测序显示12株CRKP中8株产KPC-2,1株产NDM-1,3株未检出碳青霉烯酶。将12株CRKP与大肠埃希菌J53进行接合,获得3株接合子。PFGE分型显示,12株CRKP可分为4个型3个亚型。MLST结果显示,8株blaKPC阳性肺炎克雷伯菌均为ST11型,1株blaNDM-1阳性肺炎克雷伯菌为ST278,3株不产碳青霉烯酶菌分别为ST76、ST37、ST610。结论产KPC-2型碳青霉烯酶是该院分离的肺炎克雷伯菌对碳青霉烯类抗菌药物耐药的主要机制,产NDM-1阳性的肺炎克雷伯菌已在该院出现。
[Abstract]:Objective to study the epidemic characteristics and drug resistance mechanism of carbapenems resistant Klebsiella pneumoniae (CRKP) isolated in children. Methods 12 carbapenems resistant Klebsiella pneumoniae isolated from clinical microorganism room of Shanghai Children's Hospital in 1-12 months of 2013 were collected, and the drug resistance to commonly used antibiotics was detected by agar dilution method. Two amine four acetic acid (EDTA) synergistic test and 3'- amino benzyl boric acid (APB) synergistic test were used to perform the phenotypic analysis of carbapenem, PCR amplification and DNA sequencing to confirm the genotype of carbapenem. The conjugation test was used to detect whether the carbapenem resistance gene was located on the plasmid, the pulse field gel electrophoresis (PFGE) and the multiple point sequence classification (MLST) detection strain The results showed that 12 strains of CRKP were sensitive to polymyxin E, and the resistance rate to cephalosporins was 100%. The resistance rate to imipenem, meropenem and eopenem was 91.7%, 91.7% and 100%.PCR and DNA sequencing showed that 8 strains of KPC-2,1 strains produced in 12 CRKP strains did not detect carbapenems. 12 CRKP and Escherichia coli J5 were in J5. 3 the 3 strains of zygote.PFGE typing showed that 12 strains of CRKP could be divided into 4 types and 3 subtypes.MLST results, and 8 strains of Klebsiella pneumoniae were ST11, 1 blaNDM-1 positive Klebsiella pneumoniae were ST76, ST37, and ST610. concluded that KPC-2 type carbapenems were isolated from the hospital. The main mechanism of carbapenem resistance to Klebsiella pneumoniae is the presence of NDM-1 positive Klebsiella pneumoniae in the hospital.
【作者单位】：上海市儿童医院上海交通大学附属儿童医院检验科;复旦大学附属华山医院抗生素研究所;
【基金】：上海卫计委重点学科-传染病与卫生微生物学(15GWZK0101) 上海市卫生计生系统重要薄弱学科建设(2015ZB0203)
【分类号】：R446.5

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