儿童下呼吸道感染耐碳青霉烯类肺炎克雷伯菌耐药性与毒力基因分析  

Analysis of resistance and virulence genes of carbapenem-resistant Klebsiella pneumoniae in children with lower respiratory tract infection

作　　者：董晓彤 杨寒 李丽丽 张敏 周昕怡 张传玲 康海全 DONG Xiao-tong;YANG Han;LI Li-li;ZHANG Min;ZHOU Xin-yi;ZHANG Chuan-ling;KANG Hai-quan(Medical Technology School,Xuzhou Medical University,Xuzhou,Jiangsu 221004,China;不详)

机构地区：[1]徐州医科大学医学技术学院,江苏徐州221004 [2]徐州市儿童医院检验科,江苏徐州221002 [3]徐州医科大学附属医院检验科,江苏徐州221002

出　　处：《中华医院感染学杂志》2023年第11期1721-1725,共5页Chinese Journal of Nosocomiology

基　　金：江苏省六大人才高峰基金资助项目(WSN-091);江苏省卫健委科研基金资助项目(Z2021009);徐州市卫生健康委青年创新基金资助项目(XWKYHT20200062);徐州医科大学大学生创新基金资助项目(202010313053Y)。

摘　　要：目的探究临床患儿分离的耐碳青霉烯类肺炎克雷伯菌(CRKP)的耐药性和毒力分子特征,助力儿童感染CRKP的治疗与防控。方法收集徐州医科大学附属医院和徐州市儿童医院2018年9月-2020年12月临床住院患儿分离的非重复的CRKP菌株62株,采用琼脂稀释法对最小抑菌浓度(MIC)进行检测,聚合酶链式反应(PCR)扩增常见的碳青霉烯类耐药基因(KPC,SME,GES,VIM,IMP,NDM,OXA-48)及12个毒力基因(rmpA、rmpA2、kfu、aerobactin、iroN、entB、ybtS、fimH、mrkD、allS、iucA、magA),对全部菌株进行wzi基因测序检测荚膜血清型,同时利用多位点测序(MLST)分析菌株间的同源性。结果62株CRKP均为多药耐药菌,对头孢菌素类、哌拉西林/他唑巴坦完全耐药,对美罗培南、亚胺培南、氨曲南、阿米卡星、环丙沙星、庆大霉素耐药率均>60%,对多黏菌素和替加环素完全敏感;62株CRKP均携带碳青霉烯酶耐药基因,主要携带blaKPC-2,占77.42%(48/62);62株CRKP均携带毒力基因,以fimH、ybtS、entB为主,分别占96.77%(60/62)、77.42%(48/62)、72.58%(45/62);MLST结果显示,45株为ST11-KL47型菌株,其毒力基因携带模式以entB+ybtS+fimH和entB+ybtS+mrkD+fimH为主。结论62株临床患儿分离的CRKP主要来自重症下呼吸道感染,血清型以ST11-KL47型为主,均为多药耐药菌,同时含有丰富的荚膜多糖、携带大量的毒力基因。因此,应加强预防措施并实施长期监测来控制高毒力高耐药肺炎克雷伯菌在儿童间的传播流行。OBJECTIVE To investigate the resistance and virulence molecular characteristics of Carbapenem-resistant Klebsiella pneumoniae(CRKP)isolated from clinical children,and to contribute to the treatment and prevention,and control of CRKP infection in children.METHODS Sixty-two non-replicated CRKP strains isolated from clinically hospitalized children in the Affiliated Hospital of Xuzhou Medical University and Xuzhou Children's Hospital from Sep.2018 to Dec.2020 were collected and tested for minimum inhibitory concentration(MIC)by agar dilution method.Common carbapenem resistance genes(blaKPC,blaSME,blaGES,blaVIM,blaIMP,blaNDM)and 12 virulence genes(rmpA,rmpA2,kfu,aerobactin,iroN,entB,ybtS,fimH,mrkD,allS,iucA,magA)were amplified by polymerase chain reaction(PCR).Wzi gene sequencing was performed on all strains to detect podoconjugate serotypes,while multi-locus sequencing(MLST)was used to analyze the homology between strains.RESULTS All 62 CRKP strains were multi-drug resistant,fully resistant to cephalosporins,piperacillin/tazobactam,and more than 60%resistant to meropenem,imipenem,aminotrans,amikacin,ciprofloxacin,gentamicin,and fully susceptible to polymyxin and tigecycline.All 62 CRKP strains carried carbapenemase resistance genes,mainly carrying blaKPC-2,accounting for 77.42%.All 62 CRKP strains carried virulence genes,mainly fimH,ybtS,and entB,accounting for 96.77%(60/62),77.42%(48/62),and 72.58%(45/62),respectively.The MLST results showed that 45 strains were ST11-KL47 type strains,with virulence gene carriage patterns dominated by entB+ybtS+fimH and entB+ybtS+mrkD+fimH.CONCLUSION The 62 strains of Klebsiella pneumoniae isolated from clinical children were mainly from severe lower respiratory tract infections,with a predominance of serotypes ST11-KL47,all of which were multi-drug resistant and rich in podoconjugates,and carried a large number of virulence genes.Therefore,preventive measures and long-term surveillance should be strengthened to control the transmission of the highly virulent and drug-resistant Kle

关 键 词：儿童 耐碳青霉烯类肺炎克雷伯菌 耐药 毒力 

分 类 号：R563.1[医药卫生—呼吸系统]