应用基质辅助激光解析电离飞行时间质谱检测产碳青霉烯酶肠杆菌科细菌对厄他培南的水解能力

发布时间：2018-04-05 09:09

本文选题：基质辅助激光解析电离飞行时间质谱　切入点：产碳青霉烯酶　出处：《中国感染与化疗杂志》2016年05期

【摘要】：目的应用基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)检测产KPC-2、NDM-1、VIM-2、IMP-1与IMP-4型碳青霉烯酶肠杆菌科细菌水解厄他培南的能力。方法收集2009年6月-2013年12月上海交通大学医学院附属新华医院各种临床标本中分离的耐碳青霉烯类肠杆菌科细菌(CRE),用改良Hodge试验进行产碳青霉烯酶的表型筛选,用PCR扩增方法检测其碳青霉烯酶基因,并经测序确认。应用MALDI-TOF MS进行厄他培南水解预试验以确定最适厄他培南浓度及孵育时间,随后采用预试验中的最适条件,应用MALDI-TOF MS检测CRE水解厄他培南的能力。结果 108株CRE中,有102株改良Hodge试验阳性,其中90株产KPC-2、4株产NDM-1、3株产VIM-2、2株产IMP-1、2株产IMP-4、1株同时产KPC-2和IMP-4型碳青霉烯酶,且在2 h内均能水解厄他培南;另外6株CRE改良Hodge试验阴性,未检测出上述碳青霉烯酶基因,且不水解厄他培南。结论临床微生物实验室应用MALDI-TOF MS能够快速准确检测CRE水解厄他培南的能力,筛查产碳青霉烯酶肠杆菌科细菌,为临床早期合理使用碳青霉烯类抗生素提供依据。
[Abstract]:Objective to detect the ability of KPC-2NdM-1 (VIM-2) IMP-1 and IMP-4 type carbapeninase enterobacteriaceae bacteria to hydrolyze ertapenem by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS).Methods from June 2009 to December 2013, a series of clinical specimens from Xinhua Hospital affiliated to Shanghai Jiaotong University Medical College were collected, and the phenotypic screening of carbapenem producing enzymes was carried out by modified Hodge test, which was isolated from various clinical specimens of Enterobacteriaceae, Shanghai Jiaotong University.The carbapenem gene was detected by PCR and confirmed by sequencing.The optimal concentration and incubation time of etapenem were determined by using MALDI-TOF MS to determine the optimal concentration and incubation time. Then, the ability of CRE hydrolysis of ertapenem was detected by MALDI-TOF MS.Results of the 108 CRE strains, 102 were positive for modified Hodge test, 90 of which produced KPC-2P 4 strains and NDM-1 + 3 strains, and 2 strains produced IMP-1 and 2 strains produced both KPC-2 and IMP-4 carbapenem, and were able to hydrolyze ertapenem within 2 hours.The other 6 strains were negative in CRE modified Hodge test and did not detect these carbapenase genes and did not hydrolyze ertapenem.Conclusion MALDI-TOF MS in clinical microbiology laboratory can quickly and accurately detect the ability of CRE to hydrolyze ertapenem and screen Enterobacteriaceae bacteria producing carbapenem so as to provide evidence for the rational use of carbapenem antibiotics in early clinical stage.
【作者单位】：上海交通大学医学院附属新华医院检验科临床微生物室;
【分类号】：R446.5

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