MALDI-TOF MS技术在多重耐药大肠埃希菌同源性分析及自建库中的应用研究  

Application of MALDI-TOF MS technology in homology analysis and self-built library of multidrug-resistant Escherichia coli

作　　者：彭海[1] 周健武 牟晓明[1] 孙静薇[1] 巢世兰[1] 张倩 徐雯[1] PENG Hal;ZHOU Jian-wu;MU Xiao-ming;SUN Jing-wei;CHAO Shi-lan;ZHANG Qian;XU Wen(Qinghai Provincial People's Hospital,Xining,Qinghai 810007,China)

机构地区：[1]青海省人民医院医学检验科,青海西宁810007

出　　处：《中华医院感染学杂志》2018年第24期3689-3694,共6页Chinese Journal of Nosocomiology

基　　金：青海省卫计委科研基金资助项目(2016-wjzdx-01)

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)技术对多重耐药大肠埃希菌同源性分析的能力,并建立多重耐药大肠埃希菌同源性质谱分型及药敏参考数据库。方法对2016年3月-2017年3月医院632株多重耐药大肠埃希菌(ESBLs)Eco进行MALDI-TOF MS鉴定,利用MALDI-Biotyper软件进行同源性分析及自建库;使用VITEK 2.0全自动细菌鉴定及药敏分析仪进行药敏实验,用WHONET5.6软件进行药敏结果分析。结果 2016年3月-2017年3月医院临床送检的标本中共检出(ESBLs)Eco 632株,检出率35.91%,检出标本主要有痰、尿液和血液,各科室检出(ESBLs)Eco的检出率分别为泌尿外科10.27%、肾内科8.69%、老年病科7.11%、急诊外科5.21%、血液风湿科4.90%;632株(ESBLs)Eco经MALDI-TOF MS同源性聚类分析,分为质谱数据差异性较大的两大簇六个分型,其中Ⅰa型27.00%、Ⅰb1型11.00%、Ⅰb2型9.00%、Ⅱa1型4.50%、Ⅱa2型33.5%、Ⅱb型15.00%;whonet5.6软件药敏分析显示六个分型的药敏结果也存在一定的差别;普外I、普外II、血液风湿科、急诊外科五个科室(ESBLs)Eco同源性相对较高(≥50%),提示以上科室可能存在院内交叉感染;初步建立了(ESBLs)Eco同源性质谱分型及药敏参考数据库并进行验证,质谱鉴定合格率83.3%;药敏比对合格率76.0%。结论 MALDI-TOF MS同源性聚类分析快速、准确,建立了(ESBLs)Eco同源性质谱分型及药敏参考数据库,大大缩短了实验室多重耐药菌的鉴定及同源性分析时间,并且在菌株鉴定的同时为临床提供药敏结果参考,指导临床第一时间经验用药,为临床多重耐药菌感染的快速诊断及治疗赢得宝贵时间。OBJECTIVE To evaluate the ability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS)in the analysis of multidrug-resistant E.coli,and to establish the mass spectrometry classification for homology and the drug sensitivity reference database of multidrug-resistant E.coli.METHODS MALDI-TOF MS was used to identify 632 strains of Escherichia coli producing extended spectrum beta lactamases(ESBLs)from Mar.2016 to Mar.2017,MALDI-Biotyper software was used for homology analysis and self-built library.VITEK 2.0 automatic bacterial identification and drug sensitivity test system was used for drug susceptibility testing,using WHONET5.6 software for drug susceptibility analysis.RESULTS A total of 632 strains of E.coli producing extended spectrum beta lactamases(ESBLs)were detected in the clinical specimens in our hospital from Mar.2016 to Mar.2017.The detection rate was 35.91%,and the specimens detected with ESBLS Eco were phlegm,urine and blood.The detection rate of ESBLS Eco was 10.27% for urology,8.69% for nephrology,7.11%for geriatrics,5.21% for emergency surgery,and 4.90% for blood rheumatology.Based on the homology clustering analysis,the 632 ESBLs-producing strains of E.coli were classified into two clusters and6 subtypes with relatively large difference in mass spectra,namely,Ⅰa(27.00%),Ⅰb1(11.00%),Ⅰb2(9.00%),Ⅱa1(4.50%),Ⅱa2(33.5%),Ⅱb(15.00%).Analysis of drug sensitivity with whonet5.6 software showed there was also difference in the drug sensitivity in the six subtypes.The homology of ESBLs-producing E.coli was relatively high(≥50%)among SurgicalⅠ,SurgicalⅡ,Blood Rheumatology and Emergency Department,suggesting that there may be nosocomial cross-infection in the above departments.The homology and drug sensitivity reference database of ESBLs-producing E.coli strains were established and validated.The qualified rate of mass spectrometry was 83.3%,and the drug susceptibility was 76.0%.CONCLUSION The homology clustering analysis by MALDI-TOF MS was rapid and a

关 键 词：MALDI-TOF MS 多药耐药 大肠埃希菌 同源性 

分 类 号：R446.5[医药卫生—诊断学]