MALDI-TOF MS在碳青霉烯耐药肺炎克雷伯菌流行病学分析中的应用
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摘要
目的:用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization-time of flight mass spectrometry,MALDI-TOF MS)检测碳青霉烯耐药的肺炎克雷伯菌(carbapenem-resistant Klebsiella pneumoniae,CRKp)同源性,并与脉冲场凝胶电泳(pulsed-field gel electrophoresis,PFGE)和多位点序列分型(multilocus sequence typing,MLST)两种检测方法进行对比,探讨质谱在日常工作中的应用价值。方法:选取2017年10—12月从河南科技大学第一附属医院院患者分离的以及ICU环境采集的碳青霉烯耐药肺炎克雷伯菌10株,检测8种耐药基因:blaTEM、blaSHV、blaCTX-M、blaKPC、blaIMP、blaNDM、blaVIM、blaOXA。采用PFGE、MLST和MALDI-TOF MS 3种方法对这些菌株进行同源性分析。结果:PFGE证实10株CRKp均为同一克隆株的传播,ST型同属于ST11。MALDI-TOF MS经过对细菌进行聚类分析和主成分分析可以区分不同型别肺炎克雷伯菌,对同型肺炎克雷伯菌(Klebsiella pneumonia,Kp)的同源性检测在80%左右。结论:MALDI-TOF MS在肺炎克雷伯菌的同源性检测方面可以应用于院内爆发局部感染的监测,与PFGE和MLST两种方法检测结果一致,但需要尽快建立规范的判断标准。
Objective:Matrix-assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS)was performed to detect the homology of carbapenem-resistant Klebsiella pneumoniae(CRKp). Compared with pulsed field gel electrophoresis(PFGE)and multilocus sequence typing(MLST),the application value of mass spectrometry in daily work was discussed. Methods:Ten strains of Klebsiella pneumoniae isolated from patients in our hospital from October 2017 to December 2017 and collected from ICU environment were collected. Eight resistant genes(blaTEM,blaSHV,blaCTX-M,blaKPC,blaIMP,blaNDM,blaVIM,and blaOXA)were detected. The homology of these strains was analyzed by PFGE,MLST and mass spectrometry(MALDI-TOF MS),and the results were compared.Results:It was confirmed that 10 strains of CRKp were all of the same cloned strains by PFGE,the ST-type of ST11. MALDI-TOF MS could be distinguished from different strains of Klebsiella pneumoniae by cluster analysis and principal component analysis. The homology of the same type of Klebsiella pneumoniae was about 80%. Conclusion:MALDI-TOF MS can be used in the detection of the homology of Klebsiella pneumoniae in nosocomial local infection,which is consistent with the results of PFGE and MLST. But it is necessary to establish the standard judgment standard as soon as possible.
Objective:Matrix-assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS)was performed to detect the homology of carbapenem-resistant Klebsiella pneumoniae(CRKp). Compared with pulsed field gel electrophoresis(PFGE)and multilocus sequence typing(MLST),the application value of mass spectrometry in daily work was discussed. Methods:Ten strains of Klebsiella pneumoniae isolated from patients in our hospital from October 2017 to December 2017 and collected from ICU environment were collected. Eight resistant genes(blaTEM,blaSHV,blaCTX-M,blaKPC,blaIMP,blaNDM,blaVIM,and blaOXA)were detected. The homology of these strains was analyzed by PFGE,MLST and mass spectrometry(MALDI-TOF MS),and the results were compared.Results:It was confirmed that 10 strains of CRKp were all of the same cloned strains by PFGE,the ST-type of ST11. MALDI-TOF MS could be distinguished from different strains of Klebsiella pneumoniae by cluster analysis and principal component analysis. The homology of the same type of Klebsiella pneumoniae was about 80%. Conclusion:MALDI-TOF MS can be used in the detection of the homology of Klebsiella pneumoniae in nosocomial local infection,which is consistent with the results of PFGE and MLST. But it is necessary to establish the standard judgment standard as soon as possible.
引文
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[11] Bernaschi P,Del Chierico F,Petrucca A,et al. Microbial tracking of multidrug-resistant Klebsiella pneumoniae isolates in a pediatric hospital setting[J]. Int J Immunopathol Pharmacol,2013,26(2):463-472
[12] Bader O. MALDI-TOF-MS-based species identification and typing approaches in medical mycology[J]. Proteomics,2013,13(5):788-799
[13] Jadhav S,Bhave M,Palombo EA. Methods used for the detection and subtyping of Listeria monocytogenes[J]. J Microbiol Methods,2012,88(3):327-341
[14] Woltes M,Rohde H,Maier T,et al. MALDI-TOF MS fingerprinting allows for discrimination of major methicillinresistant Staphylococcus aureus lineages[J]. Int J Med Microbiol,2011,301(1):64-68
[15] Sachse S,Bresan S,Erhard M,et al. Comparison of multilocus sequence typing,RAPD,and MALDI-TOF mass spectrometry for typing ofβ-lactam-resistant Klebsiella pneumoniae strains[J]. Diagn Microbiol Infect Dis,2014,80(4):267-271
[16]Mencacci A,Monari C,Leli C,et al. Typing of nosocomial outbreaks of Acinetobacter baumannii by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry[J]. J Clin Microbiol,2013,51(2):603-606
[2] Clinical and Laboratory Standards Institute. M100S. Performance standards for antimicrobial susceptibility testing:twenty-seventh edition[S]. Wayne,Pa:CLSI,2017
[3]陈金云,傅鹰,杨青,等. KPC-2及IMP-4酶介导肠杆菌科细菌碳青霉烯类耐药研究[J].中华微生物学和免疫学杂志,2015,35(6):419-426
[4] Piekarska K,Zacharczuk K,Szych J,et al. Dissemination of the KPC carbapenemase producing Klebsiella pneumoniae in a hospital in Warsaw,Poland[J]. Med Dosw Mikrobiol,2010,62(1):9-20
[5]朱佩琼,蒋琰,王燕飞,等.质粒介导产超广谱β-内酰胺酶肺炎克雷伯菌对磷霉素的耐药情况及其机制[J].中华传染病杂志,2016,34(5):288-291
[6] Tenover FC,Arbeit RD,Goering RV,et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis:criteria for bacterial strain typing[J]. J Clin Microbiol,1995,33(9):2233-2239
[7]李冬菊. MALDI-TOF MS在院内感染病原菌流行病学分析中的应用研究[D].青岛:青岛大学,2016:5-8
[8] Lee JH,Bae IK,Lee SH. New defirfitions of extendedspecmtrnβ-lactamase conferring worldwide emerging antibiotic resistance[J]. Med Res Rev,2012,32(1):216-232
[9] Olive DM,Bean P. Principles and application of methods for DNA-based typing of microbial[J]. J Clin Microbiol,1999,37(6):1661-1669
[10]姬小薇,廖亚玲,毛旭虎,等. MLST分析在病原微生物基因分型应用中的研究进展[J].国际检验医学杂志,2011,32(2):246-249
[11] Bernaschi P,Del Chierico F,Petrucca A,et al. Microbial tracking of multidrug-resistant Klebsiella pneumoniae isolates in a pediatric hospital setting[J]. Int J Immunopathol Pharmacol,2013,26(2):463-472
[12] Bader O. MALDI-TOF-MS-based species identification and typing approaches in medical mycology[J]. Proteomics,2013,13(5):788-799
[13] Jadhav S,Bhave M,Palombo EA. Methods used for the detection and subtyping of Listeria monocytogenes[J]. J Microbiol Methods,2012,88(3):327-341
[14] Woltes M,Rohde H,Maier T,et al. MALDI-TOF MS fingerprinting allows for discrimination of major methicillinresistant Staphylococcus aureus lineages[J]. Int J Med Microbiol,2011,301(1):64-68
[15] Sachse S,Bresan S,Erhard M,et al. Comparison of multilocus sequence typing,RAPD,and MALDI-TOF mass spectrometry for typing ofβ-lactam-resistant Klebsiella pneumoniae strains[J]. Diagn Microbiol Infect Dis,2014,80(4):267-271
[16]Mencacci A,Monari C,Leli C,et al. Typing of nosocomial outbreaks of Acinetobacter baumannii by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry[J]. J Clin Microbiol,2013,51(2):603-606