基质辅助激光解吸电离飞行时间质谱直接检测产碳青霉烯酶肠杆菌科细菌的临床价值评估  

Evaluation of clinical value of direct detection of carbapenemase-producing Enterobacteriaceae by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

作　　者：许春燕[1] 郭锋[2] 钱娇[1] 蔡家欣 余素飞[1] XU Chun-yan;GUO Feng;QIAN Jiao;CAI Jia-xin;YU Su-fei(Department of Clinical Laboratory,Taizhou Hospital,Taizhou,Zhejiang 317000,China)

机构地区：[1]浙江省台州医院检验科,浙江台州317000 [2]浙江省台州医院质量改进部,浙江台州317000

出　　处：《中国卫生检验杂志》2021年第19期2348-2351,2356,共5页Chinese Journal of Health Laboratory Technology

基　　金：2019年度台州恩泽医疗中心(集团)科研基金一般课题项目B类(19EZB14)。

摘　　要：目的 探讨基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)直接检测产碳青霉烯酶肠杆菌科细菌的临床价值。方法 以2017年3月-2020年3月本院分离的80株产碳青霉烯酶肠杆菌科细菌(CPE)为研究对象,通过PCR检测各菌株携带碳青霉烯酶基因,同时将菌株与0. 25 mg/ml亚胺培南溶液孵育60 min后离心,MALDI-TOF MS检测上清,通过菌株水解药物出现特征峰来判断菌株是否产碳青霉烯酶,并与PCR结果进行比较,同时以同期分离不产碳青霉烯酶肠杆菌科细菌(NCPE)作阴性对照。结果 80株菌株均检测出一种或多种碳青霉烯酶基因。0. 25 mg/ml亚胺培南溶液与1 ml 4麦氏浊度的菌悬液混匀后37℃孵育60 min,离心取上清进行MALDI-TOF MS检测,亚胺培南所对应的质谱峰(300±0. 65) m/z完全消失,表明该菌株产碳青霉烯酶,与PCR结果高度一致。结论 MALDI-TOF MS可以通过检测抗生素特征峰的变化来快速检测CPE,为临床快速诊断病原菌提供依据。Objective To explore the clinical value of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS)in the direct detection of carbapenemase-producing Enterobacteriaceae.Methods Eighty strains of carbapenemase-producing Enterobacteriaceae(CPE)isolated from March 2017 to Marell 2020 in our hospital were used as the research objects.PCR was used to detect the carbapenemase gene of each strain,and after incubating with 0.25 mg/ml imipenem solution for 60 min centrifugation,the supernatant was detected by MALDI-TOF MS,and the characteristic peak of the strain hydrolyzed drug was used to determine whether the strain produced carbapenemase and compared with the PCR result.In the same period,the carbapenemase-non-producing Enterobacteriaceae(NCPE)strains were isolated as negative control.Results One or rnorr carbapenemase genes were detected from 80 strains.After 0.25 mg/ml imipenem solution was mixed with 1 ml 4 McGrilh turbidity suspension,the supernatant was incubated at 37℃.for 60 min and centrifugated for MALDI-TOF MS detection.The mass spectrum peak(300±0.65)m/z corresponding to imipenem disappeared completely,indicating that the strain produced carbapenase.It was highly consistent with PCR results.Conclusion MALDI-TOF MS can quickly detect CPE by detecting changes in the characteristic peaks of antibiotics,and provide basis for rapid clinical diagnosis of pathogens.

关 键 词：基质辅助激光解吸电离 肠杆菌科细菌 碳青霉烯酶 耐药 

分 类 号：R446.5[医药卫生—诊断学]