基于国产质谱Autof ms1000快速检测耐β-内酰胺类抗菌药物肠杆菌的研究评价  

Research and evaluation of rapid detection of β-lactam-resistant enterobacteria based on domestic mass spectrometry Autof ms1000

作　　者：李海军[1] 边沁 何先伟[1] 张韩[1] 冯金芳 LI Haijun;BIAN Qin;HE Xianwei;ZHANG Han;FENG Jinfang(Department of Clinical Laboratory,Guangyuan Central Hospital,Guangyuan,Sichuan 628000,China)

机构地区：[1]广元市中心医院检验科,四川广元628000

出　　处：《国际检验医学杂志》2020年第17期2104-2111,共8页International Journal of Laboratory Medicine

摘　　要：目的建立一种基于基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)快速检测不同肠杆菌耐药性的方法,对比MALDI-TOF MS检测结果与常规药敏检测结果,评估MALDI-TOF MS在检测细菌耐药性方面的可行性。方法收集10株肠杆菌科菌株进行预实验(8株耐药,2株敏感质控),用β-内酰胺类抗菌药物(氨苄西林、亚胺培南、美罗培南、厄他培南、头孢噻肟、头孢他啶)孵育耐药细菌一定时间后,耐药菌会引起抗菌药物的水解,从而造成+18的质量偏移,通过MALDI-TOF MS检测药物水解情况,得到细菌耐药性与敏感性分类。同时用微量肉汤稀释法进行药敏实验,对比两种方法的药敏结果。采用预实验中的最佳实验条件,选择120株肠杆菌科菌株进行批量验证实验,对两种结果进行分析统计。结果抗菌药物与耐药菌孵育4 h内,氨苄西林水解在相对分子质量为350、372、394处峰消失,亚胺培南在相对分子质量为300处峰消失、美罗培南在相对分子质量为384、406处峰消失,厄他培南在相对分子质量为475、498、520处峰消失,头孢噻肟在相对分子质量为456、478处峰消失,头孢他啶在相对分子质量为468处峰消失,敏感菌株特征峰仍然存在,由此可判断菌株的耐药性。批量验证结果与微量肉汤稀释法结果一致,且可在4 h内得到药敏结果。结论国产质谱Autof ms1000可用于快速检测β-内酰胺类细菌耐药情况,结果准确可靠,与常规药敏实验方法符合率高,并且缩短了检测时间,可提前指导临床早期合理使用β-内酰胺类抗菌药物。Objective To establish a method based on matrix-assisted laser disorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS)for rapid detection of drug resistance of different enterobacteria,to compare the results of MALDI-TOF MS with the results of conventional susceptibility testing to evaluate the feasibility of MALDI-TOF MS in detecting bacterial resistance.Methods A total of 10 strains of Enterobacteriaceae were collected for pre-experiment(8 resistant strains,2 sensitive quality control strains).After incubating resistant bacteria withβ-lactam antibacterial drugs(ampicillin,imipenem,meropenem,ertapenem,cefotaxime,ceftazidime)for a certain period of time.Drug-resistant bacteria could cause the hydrolysis of antibacterial drugs,resulting in a mass shift of+18.The drug hydrolysis was detected by MALDI-TOF MS to obtain the classification of bacterial resistance and sensitivity.At the same time,the susceptibility test was carried out with the micro broth dilution method to compare the susceptibility results of the two methods.Using the best experimental conditions in the preliminary experiment,120 strains of Enterobacteriaceae were selected for batch verification experiments,and the two results were analyzed and counted.Results Within 4 hours of incubation between antimicrobial drugs and drug-resistant bacteria,ampicillin was hydrolyzed,and the peaks at relative molecular masses of 350,372,and 394 disappeared,and the peaks at relative molecular masses of 300 disappeared for imipenem.The peaks at relative molecular masses of 384 and 406,meropenem disappeared.The peaks at relative molecular masses of 475,498,and 520,ertapenem disappeared.The peaks at the relative molecular masses of 456 and 478,cefotaxime disappeared.The peak of ceftazidime at the relative molecular weight of 468 disappeared,and the characteristic peak of sensitive strains still existed,so the drug resistance of the strain could be judged.The batch verification results were consistent with the results of the micro broth dilution method,and

关 键 词：基质辅助激光解吸电离飞行时间质谱 Β-内酰胺类抗菌药物 肠杆菌科细菌 耐药性检测 

分 类 号：R372[医药卫生—病原生物学]