MALDI-TOF MS用于碳青霉烯耐药肺炎克雷伯菌同源性分析的初步研究  

Homology analysis of Carbapenems-resistant klebsiella pneumoniae by MALDI-TOF MS

作　　者：张仁峰 张炳昌[1] 邵春红[1] 范会[1] 王丽萍[1] 金炎[1] Zhang Renfeng;Zhang Bingchang;Shao Chunhong;Fan Hui;Wang Liping;Jin Yan(Department of clinicial laboratory,Shandong Provincial Hospital affiliated to Shandong University,Jinan 250021,China)

机构地区：[1]山东大学附属山东省立医院检验科,济南250021

出　　处：《中华检验医学杂志》2018年第8期589-595,共7页Chinese Journal of Laboratory Medicine

基　　金：山东省重点研发计划（2016GSF201078）

摘　　要：目的 评价基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）技术对碳青霉烯耐药肺炎克雷伯菌（CRKPN）同源性分析的能力.方法 收集山东大学附属省立医院儿科病房及心外科病房2011年4月至2013年10月分离的21株CRKPN菌株,分别采用脉冲场凝胶电泳（PFGE）、多位点序列分型（MLST）及MALDI-TOF MS技术进行回顾性同源性分析.结果 MALDI-TOF MS将21株CRKPN菌株根据亲缘关系远近分为三群,其中17例菌株为II群,同源性高于75%,从获得的17株菌的蛋白质谱图结果分析,其蛋白峰大致一致,由此推断其亲缘关系比较接近,与PFGE和MLST结果基本一致.而同源性较低（〈60%）的H13与上述菌株存在差异,尤其在分子量4365、5381及6289处蛋白峰有显著差异,PFGE分析显示H13与其他菌株的同源性为61.0%,MLST分型为ST54.结论 MALDI-TOF MS技术能够准确对碳青霉烯耐药肺炎克雷伯菌进行同源性分析,较之其他同源性分析方法,快速方便,可满足医院感染工作的需求,及时防止耐药菌株在医院感染的爆发与流行.Objective To evaluate the ability of matrix-assisted laser desorption/ionization-time of flight mass spectrometry （ MALDI-TOF MS ） in the homology analysis of Carbapenems-resistant klebsiella pneumonia. Methods Twenty-one non-duplicated strains of Carbapenems-resistant klebsiella pneumoniae were isolated from Shandong Provincial Hospital affiliated to Shandong University during April 2011 and October 2013 in this study. Twenty isolates were from neonatal unit and one from cardiac surgery. The homology analysis of Carbapenems-resistant klebsiella pneumoniae was performed with pulsed-field gel electrophoresis （PFGE）, muhiloeus sequence typing （MLST） and MALDI-TOF MS respectively. Results The result of PFGE was consistent with MLST. The twenty-one CRKPN strains were divided into three groups by MALDI-TOF MS according to their relationship, 18 of them belonged to II group, and the homology was higher than 75%. From the analysis of protein mass spectra of 18 strains, the protein peaks were roughly the same. Thus, it was concluded that their relationship was close, and the results were basically consistent with the results of PFGE and MLST. The H13 strain with low homology （ 〈 60% ） was different from the above strains, especially in the molecular weight 4365, 5381 and 6289. The PFGE analysis showed that the homology between H13 and other strains was 61%, and the MLST classification result was ST54. Concinsions MALDI-TOF MS can be used to identify CRKPN accurately and analyze its homology analysis more conveniently than other methods in clinical laboratory. MALDI-TOF MS has the potential to be used as an easy and rapid epidemiology typing tool for nosocomial infection investigation caused by drug-resistant bacteria.

关 键 词：肺炎克雷伯菌 碳青霉烯类 抗药性 细菌 序列同源性 光谱法 质量 基质辅助激光解吸电离 

分 类 号：R446.5[医药卫生—诊断学]