基质辅助激光解吸电离飞行时间质谱用于致病性弧菌鉴定
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摘要
采用致病性弧菌标准菌株和分离菌株,在规定的培养条件和检测方法下进行MALDI-TOFMS检测,同时进行方法的重复性、稳定性和特异性分析。新增加致病性弧菌菌株包括12种致病性弧菌,菌株来源广泛,构建的致病性弧菌MALDI-TOF-MS数据库信息量大、特异性强。结果表明,MALDITOF-MS方法可以快速、准确地鉴定致病性弧菌,可以用于致病性弧菌的日常检测鉴定。
Standard strains and isolated strains of pathogenic vibrio were used for MALDI-TOF-MS detection under specified culture conditions and detection methods. Meanwhile,the repeatability,stability and specificity of the method were analyzed to confirm the accuracy of the method. With main pathogenic vibrio from various sources,the strain database was complete with high specificity. The results show that MALDI-TOF-MS can be used to identify the pathogenic vibrio rapidly and accurately.
Standard strains and isolated strains of pathogenic vibrio were used for MALDI-TOF-MS detection under specified culture conditions and detection methods. Meanwhile,the repeatability,stability and specificity of the method were analyzed to confirm the accuracy of the method. With main pathogenic vibrio from various sources,the strain database was complete with high specificity. The results show that MALDI-TOF-MS can be used to identify the pathogenic vibrio rapidly and accurately.
引文
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[2] FARMER J J,3RD J M,BIRKHEAD K. Vibrio[M]∥Murray P R,Baron E J,JORGENSEN J H. Manual of clinical microbiology. Washington D C:ASM Press,2003:706-718.
[3] ERLER R,WICHELS A,HEINEMEYER E A. Vibrio Base:a MALDI-TOF MS database for fast identification of Vibrio spp. that are potentially pathogenic in humans[J]. Systematic and Applied Microbiology,2015,38(1):16-25.
[4] LETCHUMANAN V,CHAN K G,LEE L H. Vibrio parahaemolyticus:a review on the pathogenesis,prevalence,and advance molecular identification techniques.[J].Front Microbiol.,2014,5(5):1-13.
[5] AKYAR I. Comprehensive analytical chemistry:food pathogens,68[M]. Netherland:Elsevier,2015:617-652.
[6] BONNIN-JUSSERAND M,COPIN S,LE BRIS C. Vibrio species involved in seafood-borne outbreaks(Vibrio cholerae,V. parahaemolyticus and V. vulnificus):Review of microbiological versus recent molecular detection methods in seafood products[J]. Critical Reviews in Food Science and Nutrition,2019,59(4):597-610.
[7] CHEN J,ZHANG R,QI X. Epidemiology of foodborne disease outbreaks caused by Vibrio parahaemolyticus during2010-2014 in Zhejiang Province,China[J]. Food Control,2017,77(2017):110-115.
[8] KIM K I,WON K M,LEE E S. Detection of vibrio and ten vibrio species in cage-cultured fish by multiplex polymerase chain reaction using house-keeping genes[J].Aquaculture,2019,506(2019):417-423.
[9] XU Y G,SUN L M,WANG Y S. Simultaneous detection of Vibrio cholerae,Vibrio alginolyticus,Vibrio parahaemolyticus and Vibrio vulnificus in seafood using dual priming oligonucleotide(DPO)system-based multiplex PCR assay[J]. Food Control,2017,71(2017):64-70.
[10] FERN NDEZ-LVAREZ C,GIJ N D,LVAREZ M,et al. First isolation of Aeromonas salmonicida subsp. salmonicida from diseased sea bass,Dicentrarchus labrax(L.),cultured in Spain[J]. Aquaculture Reports,2016,4(2016):36-41.
[11] FERN NDEZ-LVAREZ C,TORRES-CORRAL Y,SALTOS-ROSERO N. MALDI-TOF mass spectrometry for rapid differentiation of Tenacibaculum species pathogenic for fish[J]. Applied Microbiology and Biotechnology,2017,101(13):5377-5390.
[12] KITATSUKAMOTO K. Phylogenetic relationships of marine-bacteria,mainly members of the family Vibrionaceae,determined on the basis of 16S rRNA sequences[J]. Int. J. Syst. Bacteriol,1993,43(1):8-19.
[13] MELLMANN A J M. MALDI-TOF mass spectrometrybased microbial identification and beyond[M]. Germany:Springer-Verlag,2018:211-234.
[14] LI P,XIN W,XIA S. MALDI-TOF mass spectrometry-based serotyping of V. parahaemolyticus isolated from the Zhejiang province of China[J]. BMC Microbiology,2018,18(1):185.
[15] CHAIKA S O,TELESMANICH N R,LOMOV Y M.MALDI-TOF mass spectrometry identification of the marker of vibrio cholerae toxicity[J]. Molecular Genetics,Microbiology and Virology,2018,33(2):145-146.
[16] SAUGET M,VALOT B,BERTRAND X. Can MALDITOF mass spectrometry reasonably type bacteria?[J].Trends in Microbiology,2017,25(6):447-455.
[17] BURBICK C R,NYDAM S D,HENDRIX G K. Use of matrix-assisted laser desorption ionization time-offlight mass spectrometry for the identification of pathogenic vibrio in fish[J]. Journal of Aquatic Animal Health,2018,30(4):332-338.
[18] HUBER I,PAVLOVIC M,MAGGIPINTO M. Interlaboratory proficiency test using MALDI-TOF MS for identification of food-associated bacteria[J]. Food Analytical Methods,2018,11(4):1068-1075.
[19] DIECKMANN R,STRAUCH E,ALTER T. Rapid identification and characterization of Vibrio species using whole‐cell MALDI‐TOF mass spectrometry[J].Journal of Applied Microbiolog,2010,109(1):199-211.
[20]丁进亚.基质辅助激光解析电离飞行时间质谱应用于临床微生物快速鉴定诊断[J].华南国防医学杂志,2018,32(4):286-299.
[21] PAAUW A,TRIP H,NIEMCEWICZ M. Omp U as a biomarker for rapid discrimination between toxigenic and epidemic Vibrio cholerae O1/O139 and non-epidemic Vibrio cholerae in a modified MALDI-TOF MS assay[J].BMC Microbiology,2014,14(1):158.
[22] TELESMANICH N R,CHA KA S O,VODIANITSKAIA S I. The application of mass spectrometry technique MALDI-TOF for inter-specific differentiation of closely-related vibrio[J]. Klinicheskaia Laboratornaia Diagnostika,2014,59(8):27-28.