MALDI-TOF MS与16S rDNA方法对弧菌科微生物的鉴定与系统分类学分析
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摘要
目的比较基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)与16S rDNA方法对弧菌科微生物的鉴定及系统分类学分析能力。方法对19株弧菌科微生物,采用MALDI-TOF MS进行蛋白质图谱采集,通过对特征峰的分析,实现对微生物的鉴定和系统分类学分析;同时对19株微生物进行16S rDNA测序,用邻接法对16S rDNA序列进行鉴定和系统分类学分析,比较两种方法在弧菌科微生物鉴定和系统分类学分析中的异同。结果两种方法对19株弧菌科微生物的种属鉴定结果一致。系统分类学分析中,多株同种属的弧菌的两种方法分析结果一致,但对拟态弧菌和霍乱弧菌在树状图中的位置和亲缘关系,两种方法差异较大。结论 MALDI-TOF MS与16S rDNA均能够快速准确地鉴定弧菌科微生物,但利用MALDI-TOF MS进行系统分类学分析还有待数据库的扩大及算法的优化。
Objective To compare the performances of matrix assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS) vs 16S rDNA for the identification and systematic taxonomic analysis of Vibrio microorganisms. Methods 19 strains of Vibrio microorganisms were detected using MALDI-TOF MS to collect protein profiles. Microbial identification and systematic taxonomic analysis were carried out to compare and analyze the characteristic peaks. Meanwhile, the 19 Vibrio microorganisms were sequenced by using 16S rDNA for identification on the level of molecular biology, and phylogenetic tree analysis of the 16S rDNA sequences was realized with Neighbor-joining method. The similarities and differences between the two methods in the identification and systematic taxonomic analysis of Vibrio microorganisms were compared. Results The results of species identification by the two methods were consistent. Systematic taxonomic analysis showed that for the Vibrio strains of the same species, the results of the two methods were consistent, however, for the locations of and relationship between Vibrio mimius and Vibrio cholerae in the dendrograms, there were differences between the two methods. Conclusion Both MALDI-TOF MS and 16S rDNA can identify Vibrio microorganisms quickly and accurately, however, the system taxonomic analysis using MALDI-TOF MS has yet to expand the database and optimize the algorithm.
Objective To compare the performances of matrix assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS) vs 16S rDNA for the identification and systematic taxonomic analysis of Vibrio microorganisms. Methods 19 strains of Vibrio microorganisms were detected using MALDI-TOF MS to collect protein profiles. Microbial identification and systematic taxonomic analysis were carried out to compare and analyze the characteristic peaks. Meanwhile, the 19 Vibrio microorganisms were sequenced by using 16S rDNA for identification on the level of molecular biology, and phylogenetic tree analysis of the 16S rDNA sequences was realized with Neighbor-joining method. The similarities and differences between the two methods in the identification and systematic taxonomic analysis of Vibrio microorganisms were compared. Results The results of species identification by the two methods were consistent. Systematic taxonomic analysis showed that for the Vibrio strains of the same species, the results of the two methods were consistent, however, for the locations of and relationship between Vibrio mimius and Vibrio cholerae in the dendrograms, there were differences between the two methods. Conclusion Both MALDI-TOF MS and 16S rDNA can identify Vibrio microorganisms quickly and accurately, however, the system taxonomic analysis using MALDI-TOF MS has yet to expand the database and optimize the algorithm.
引文
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[9] LI Yicai,ZHAO Feng,ZHU Lanlan,et al.Investigations on contamination and serotype of Vibrio parahaemolyticus in shellfish obtained from Yellow Sea and Bohai Sea area[J].J Southern Agric,2012,43(2):241-244.(in Chinese) 李毅财,赵峰,朱兰兰,等.黄渤海区域贝类中副溶血弧菌污染调查及血清学分型[J].南方农业学报,2012,43(2):241-244.
[10] Seng P,Drancourt M,Gouriet F,et al.Ongoing revolution in bacteriology:Routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry[J].Clin Infect Dis,2009,49(4):543-551.
[11] WEI Chao,DAI Xiaohang,GUO Lingan,et al.Analysis and comparsion of MALDI-TOF MS and 16S rDNA for the identification of Enterobacteriaceae[J].J Chin Mass Spectrometry Society,2018,39(2):209-215.(in Chinese) 魏超,代晓航,郭灵安,等.MALDI-TOF MS与16S rDNA方法对肠杆菌科微生物鉴定比较分析[J].质谱学报,2018,39(2):209-215.
[12] Krishnamurthy T,Davis MT,Stahl DC,et al.Liquid chromatography/microspray mass spectrometry for bacterial investigations[J].Rapid Commun Mass Spectrom,1999,13(1):39-49.
[2] Van Veen SQ,Claas EC,Kuijper EJ.High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization-time of flight mass spectrometry in conventional medical microbiology laboratories[J].J Clin Microbiol,2010,48(3):900-907.
[3] Bessède E,Angla-Gre M,Delagarde Y,et al.Matrix-assisted laser-desorption/ionization biotype:Experience in the routine of a University hospital[J].Clin Microbiol Infect,2011,17(4):533-538.
[4] Majcherczyk PA,McKenna T,Moreillon P,et al.The discriminatory power of MALDI-TOF mass spectrometry to differentiate between isogenic teicoplanin-susceptible and teicoplanin-resistant strains of methicillin-resistant Staphylococcus aureus[J].FEMS Microbiol Lett,2006,255(2):233-239.
[5] Stevenson LG,Drake SK,Murray PR.Rapid identification of bacteria in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry[J].J Clin Microbiol,2010,48(2):444-447.
[6] 王镜岩,朱圣庚,徐长法.生物化学[M].3版.北京:高等教育出版社,2006:522-523.
[7] LIU Chaojun,SHEN Dingxia.Application of 16S rDNA sequencing for identification of bacteria[J].J Chin PLA Postgrad Med Sch,2011,32(7):774-776.(in Chinese) 刘朝军,沈定霞.16S rDNA序列测定在细菌鉴定中的应用[J].解放军医学院学报,2011,32(7):774-776.
[8] YAN Binlun,QIN Guomin,BAO Zenghai,et al.Isolation and identification of Vibrio parahaemolyticus from diseased Portunus triuberculatus L[J].Marine Sci Bull,2010,29(5):560-566.(in Chinese) 阎斌伦,秦国民,暴增海,等.三疣梭子蟹病原副溶血弧菌的分离与鉴定[J].海洋通报,2010,29(5):560-566.
[9] LI Yicai,ZHAO Feng,ZHU Lanlan,et al.Investigations on contamination and serotype of Vibrio parahaemolyticus in shellfish obtained from Yellow Sea and Bohai Sea area[J].J Southern Agric,2012,43(2):241-244.(in Chinese) 李毅财,赵峰,朱兰兰,等.黄渤海区域贝类中副溶血弧菌污染调查及血清学分型[J].南方农业学报,2012,43(2):241-244.
[10] Seng P,Drancourt M,Gouriet F,et al.Ongoing revolution in bacteriology:Routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry[J].Clin Infect Dis,2009,49(4):543-551.
[11] WEI Chao,DAI Xiaohang,GUO Lingan,et al.Analysis and comparsion of MALDI-TOF MS and 16S rDNA for the identification of Enterobacteriaceae[J].J Chin Mass Spectrometry Society,2018,39(2):209-215.(in Chinese) 魏超,代晓航,郭灵安,等.MALDI-TOF MS与16S rDNA方法对肠杆菌科微生物鉴定比较分析[J].质谱学报,2018,39(2):209-215.
[12] Krishnamurthy T,Davis MT,Stahl DC,et al.Liquid chromatography/microspray mass spectrometry for bacterial investigations[J].Rapid Commun Mass Spectrom,1999,13(1):39-49.
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