基质辅助激光解吸电离飞行时间质谱在醋酸钙-鲍曼不动杆菌复合菌鉴定及同源性分析中的应用
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摘要
目的评价基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)对我院血流感染样本中醋酸钙-鲍曼不动杆菌复合菌的鉴定及同源性分析中的应用。方法收集2011年4月—2015年4月我院非重复血流感染标本,对VITEK 2 Compact全自动微生物鉴定仪初步鉴定为醋酸钙-鲍曼不动杆菌复合菌的菌株,开展基质辅助激光解吸电离飞行时间质谱鉴定,用MALDIBiotyper软件和ERIC-PCR进行同源性分析,并用16S r RNA及rec A基因测序技术进行验证。结果基质辅助激光解吸电离飞行时间质谱对20株VITEK 2无法鉴定到种的醋酸钙-鲍曼不动杆菌复合菌展开了有效的鉴定,其中9株为医院不动杆菌(Acinetobacter nosocominalis),11株为皮特不动杆菌(Acinetobacter pittii),两种细菌的质谱图中均含有特异性的峰图,能有效地将两类细菌进行区分。同源性分析结果将菌株分为2大簇(I型11株,II型9株),II型又分为3小簇(IIa4株,IIb3株,IIc2株)。I型、II型分布在我院4个病区。结论基质辅助激光解吸电离飞行时间质谱在醋酸钙-鲍曼不动杆菌复合菌鉴定方面具有精准、快速的优点,可以鉴别出皮特不动杆菌和医院不动杆菌,MALDI-Biotyper软件进行同源性分析十分便捷。
Objective To evaluate the application of laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) in the identification and homology analysis of Acinetobacter calcoaceticus-baumannii(Acb) complex isolated from bloodstream infections.Methods During April 2011 to April 2015,non-repetitive Acb complex strains recovered from patients with bloodstream infections were collected in our hospital.All isolates were evaluated by MALDITOF MS.The homology of Acb complex isolates was analyzed by Biotyper software and ERIC-PCR,and was confirmed by 16 S r RNA and rec A genes sequencing.Results In total,twenty Acb complex strains were collected in this study,nine isolates were identified as Acinetobacter nosocominalis and eleven isolates were Acinetobacter pittii by MALDI-TOF MS.The twenty Acb complex strains were divided into two clusters(type I and type II) by MALDI-TOF MS Biotyper,and type II was divided into three sub-clusters(type IIa,type IIb and type IIc).Conclusion All the isolates were correctly identified by MALDI-TOF MS compared to the identifications obtained by 16 S r RNA sequencing.MALDI-TOF MS enabled the rapid and reliable identification of the pathogen species belonging to the Acb complex,which indicated that it is a good alternative to molecular biology.
Objective To evaluate the application of laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) in the identification and homology analysis of Acinetobacter calcoaceticus-baumannii(Acb) complex isolated from bloodstream infections.Methods During April 2011 to April 2015,non-repetitive Acb complex strains recovered from patients with bloodstream infections were collected in our hospital.All isolates were evaluated by MALDITOF MS.The homology of Acb complex isolates was analyzed by Biotyper software and ERIC-PCR,and was confirmed by 16 S r RNA and rec A genes sequencing.Results In total,twenty Acb complex strains were collected in this study,nine isolates were identified as Acinetobacter nosocominalis and eleven isolates were Acinetobacter pittii by MALDI-TOF MS.The twenty Acb complex strains were divided into two clusters(type I and type II) by MALDI-TOF MS Biotyper,and type II was divided into three sub-clusters(type IIa,type IIb and type IIc).Conclusion All the isolates were correctly identified by MALDI-TOF MS compared to the identifications obtained by 16 S r RNA sequencing.MALDI-TOF MS enabled the rapid and reliable identification of the pathogen species belonging to the Acb complex,which indicated that it is a good alternative to molecular biology.
引文
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[9]邹自英,陈莉,刘媛,等.分子生物学方法鉴定鲍曼不动杆菌与仪器鉴定结果比较[J].四川医学,2015,35(9):1123-1125.
[10]王卫萍,奚海燕,黄梅,等.基质辅助激光解吸电离飞行时间质谱在临床病原菌鉴定中的应用[J].临床检验杂志,2015,5(33):337-340.
[11]黎敏,皮雯雯,王艺,等.醋酸钙-鲍曼不动杆菌复合菌的分子流行病学及体外药物敏感性分析[J].国际检验医学杂志,2015,36(19):2803-2805.
[2]Sahl J W,Gillece J D,Schupp J M,et al.Evolution of a pathogen:A comparative genomics analysis identifies a genetic pathway to pathogenesis in Acinetobacter[J].PLo S ONE,2013,8(1):e54287.
[3]Boo T W,Walsh F,Crowley B.Molecular characterization of carbapenem-resistant Acinetobacter species in an Irish university hospital:predominance of Acinetobacter genomic species 3[J].J Med Microbiol,2009,58(Pt 2):209-216.
[4]Wisplinghoff H,Paulus T,Lugenheim M,et al.Nosocomial bloodstream infections due to Acinetobacter baumannii,Acinetobacter pittii and Acinetobacter nosocomialis in the United States[J].J Infect,2012,64(3):282-290.
[5]Wang X,Chen T,Yu R,et al.Acinetobacter pittii and Acinetobacter nosocomialis among clinical isolates of the Acinetobacter calcoaceticus-baumannii complex in Sichuan China[J].Diagn Microbiol Infect Dis,2013,76(3):392-395.
[6]Lee N Y,Chang TC,Wu C J,et al.Clinical manifestations,antimicrobial therapy,and prognostic factors of monomicrobial Acinetobacter baumannii complex bacteremia[J].J Infect,2010,61(3):219-227.
[7]Versalovic J,Koeuth T,Lupski J R.Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes[J].Nucleic Acids Res,1991,19(24):6823-6831.
[8]陈佰义,何礼贤,胡必杰,等.中国鲍曼不动杆菌感染诊治与防控专家共识[J].中国医药科学,2012,2(8):1-8.
[9]邹自英,陈莉,刘媛,等.分子生物学方法鉴定鲍曼不动杆菌与仪器鉴定结果比较[J].四川医学,2015,35(9):1123-1125.
[10]王卫萍,奚海燕,黄梅,等.基质辅助激光解吸电离飞行时间质谱在临床病原菌鉴定中的应用[J].临床检验杂志,2015,5(33):337-340.
[11]黎敏,皮雯雯,王艺,等.醋酸钙-鲍曼不动杆菌复合菌的分子流行病学及体外药物敏感性分析[J].国际检验医学杂志,2015,36(19):2803-2805.