MALDI-TOF MS对猪源肠球菌的快速检测
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摘要
应用基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOF MS)和MALDI-Biotype 3软件对河南分离株肠球菌进行快速鉴定及同源性分析。使用甲酸提取法按照MALDI-TOF-MS要求进行样品制备,点靶并获取样品质谱图;运用Biotyper分析软件对所获取的质谱图进行鉴定和同源性分析。同时用Vitek-2全自动微生物鉴定系统进行辅助分析鉴定。结果显示,应用MALDI-TOF-MS方法鉴定的33株肠球菌中粪肠球菌13株,屎肠球菌20株,且可信度分值大于2.300,能完全鉴定到种的水平。PCA聚类分析显示,与屎肠球菌相比,粪肠球菌的亲缘关系更近一些。而Vitek-2对屎肠球菌的鉴定结果差异较大。结果表明,MALDI-TOF MS对肠球菌的快速鉴定和同源性分析,操作快速简便,准确率高,有利于兽医临床快速诊断,将成为兽医微生物常用诊断工具。
Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry(MALDI-TOF-MS)instrument and the Biotyper 3 analysis software were applied to rapid identification and homology analysis of Enterococcus isolates fromfor infected pigs in Henan.The bacteria sample were prepared by formic acid extraction method of MALDI-TOF MS then added to the target polished steel plate and the mass spectra was acquired.The spectra was analyzed by the Biotyper 3 analysis software.And automatic bacteria identification system was used to aid the analysis and identification of the Enterococcus.The MALDI-TOF MS determination showed that a total of 30 isolates were Enterococcusspecies,among them,13 E.faecalis and 20 E.faecium.Confidence score was greater than 2.300,with highly probable species identification.PCA cluster analysis showed that relationship of E.faecalis were more close than E.faecium.Indentification results of E.faecium were different by automatic bacteria identification system(Vitek-2).Using MALDI-TOF MS to identify and homology analysis of Enterococcus is fast,simple,accurate,and it can be used as commonly diagnostic tools of veterinary microbiology.
Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry(MALDI-TOF-MS)instrument and the Biotyper 3 analysis software were applied to rapid identification and homology analysis of Enterococcus isolates fromfor infected pigs in Henan.The bacteria sample were prepared by formic acid extraction method of MALDI-TOF MS then added to the target polished steel plate and the mass spectra was acquired.The spectra was analyzed by the Biotyper 3 analysis software.And automatic bacteria identification system was used to aid the analysis and identification of the Enterococcus.The MALDI-TOF MS determination showed that a total of 30 isolates were Enterococcusspecies,among them,13 E.faecalis and 20 E.faecium.Confidence score was greater than 2.300,with highly probable species identification.PCA cluster analysis showed that relationship of E.faecalis were more close than E.faecium.Indentification results of E.faecium were different by automatic bacteria identification system(Vitek-2).Using MALDI-TOF MS to identify and homology analysis of Enterococcus is fast,simple,accurate,and it can be used as commonly diagnostic tools of veterinary microbiology.
引文
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[10]王耀,曹际娟,赵昕,等.单增李斯特氏菌MALDITOF-MS鉴定与分型研究[J].食品科学,2012,33(3):194-198.
[11]DALLAGASSA C B,HUERGO L F,STETS M I,et al.Matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis of Escherichia coli categories[J].Genet Mol Res,2014,13(1):716-722.
[2]WERTH B J,STEED M E,IRELAND C E,et al.Defining daptomycin resistance prevention exposures in vancomycin-resistant Enterococcus faeciumand E.faecalis[J].Antimicrob Agents Chemother,2014,58(9):5253-5261.
[3]IRANKHAH S,SOUDI M R,GHARAVI S.Ex situ study of Enterococcus faecalis survival in the recreational waters of the southern coast of the Caspian Sea[J].Iran J Microbiol,2016,8(2):101-107.
[4]王文,张玉英,李刚,等.肠球菌对氟喹诺酮类药物的敏感性及其主动外排机制的研究[J].中华检验医学杂志,2014,37(4):298-301.
[5]HUDSON C R,FEDORKA-CRAY P J,JACKSONHALL M C,et al.Anomalies in species identification of enterococci from veterinary sources using a commercial biochemical identification system[J].Lett Appl Microbiol,2003,36(4):245-250.
[6]THERESA M,TORUNN P,ROB W,et al.Investigating the mobilome in clinically important lineages of nterococcus faecium and Enterococcus faecalis[J].BMC Genomics,2015,16:282.
[7]周千渝,张延国,黄成才,等.食源性致病菌MALDITOF MS检测方法的建立与应用[J].食品工业科技,2015,36(18):59-64.
[8]战晓微,傅俊范,郑秋月,等.沙门菌MALDI-TOF MS检测方法的建立[J].现代食品科技,2011,27(5):595-597.
[9]鲍春梅,崔恩博,陈鹏,等.MALDI-TOF-MS鉴定宋内志贺菌的初步应用研究[J].传染病信息,2012,25(1):10-13.
[10]王耀,曹际娟,赵昕,等.单增李斯特氏菌MALDITOF-MS鉴定与分型研究[J].食品科学,2012,33(3):194-198.
[11]DALLAGASSA C B,HUERGO L F,STETS M I,et al.Matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis of Escherichia coli categories[J].Genet Mol Res,2014,13(1):716-722.