采用基质辅助激光解析电离飞行时间质谱快速检测产KPC大肠埃希菌ST131和ST405分型的研究

发布时间：2018-04-02 22:04

本文选题：基质辅助激光解析电离飞行时间质谱　切入点：大肠埃希菌　出处：《中国感染与化疗杂志》2016年04期

【摘要】：目的采用基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)快速检测产KPC大肠埃希菌ST131、ST405,并对该方法的可行性进行评价。方法收集浙江省人民医院临床分离46株大肠埃希菌,扩增blaKPC耐药基因,采用多位点序列分型方法(MLST)进行分型。MALDI-TOF MS对大肠埃希菌进行鉴定并对不同克隆的峰图进行主成分分析和算法统计,获得分型区分的特征峰。结果 46株KPC型大肠埃希菌均携带KPC-2耐药基因,MLST方法检测到2种ST型,分别为ST131和ST405。主成分分析图显示ST131和ST405可分为2簇,其中2株ST131和3株ST405分类错误。ClinProTools软件4种算法结果基本相似,方法特异度和灵敏度分别为93.27%和97.92%。用于区分ST131和ST405的特征峰主要为8 331.84、4 166.44、4 860.21和2 783.66 Da。这4个峰区分具有统计学意义,有较高准确性。结论采用MALDI-TOF MS可快速区分产KPC大肠埃希菌ST131和ST405,具有操作简单、耗时短、区分灵敏度和特异度高以及成本低等优点,能用于大肠埃希菌克隆分型研究。
[Abstract]:Objective to detect KPC producing Escherichia coli ST131 and ST405 by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF), and to evaluate the feasibility of this method.Methods 46 clinical isolates of Escherichia coli were collected from Zhejiang Provincial people's Hospital and blaKPC resistance genes were amplified.Multilocus sequence typing method was used to identify Escherichia coli by MALDI-TOF MS. Principal component analysis (PCA) and algorithm statistics were performed on different cloned peaks to obtain characteristic peaks.Results 46 strains of KPC Escherichia coli were detected by MLST with KPC-2 resistance gene. The results were ST131 and ST405, respectively.Principal component analysis (PCA) showed that ST131 and ST405 could be divided into 2 clusters, among which 2 strains of ST131 and 3 strains of ST405 were classified wrong. The results of the four algorithms were similar. The specificity and sensitivity of the methods were 93.27% and 97.92%, respectively.The main characteristic peaks used to distinguish ST131 from ST405 are 8 331.84 4 166.44 Li 4 860.21 and 2 783.66 Da.These four peaks are statistically significant and have high accuracy.Conclusion MALDI-TOF MS can be used to distinguish KPC producing Escherichia coli ST131 and ST405 rapidly. It has the advantages of simple operation, short time consuming, high sensitivity, high specificity and low cost. It can be used for the cloning and typing of Escherichia coli.
【作者单位】：浙江大学附属第二医院检验科;浙江省人民医院;
【基金】：浙江省中医药管理局资助项目(2011ZA058) 浙江省科学技术厅2014年度省公益性技术应用研究计划项目(2014C33191) 浙江省科技厅公共卫生监测与突发事件处置关键技术研究科技创新团队项目(2011R50021)
【分类号】：R440

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