目的 评价16S核糖体RNA基因(16S rRNA)、RNA聚合酶β亚基基因(rpoB)及基质辅助激光解析/电离飞行时间质谱(MALDI-TOF MS)技术对海洋副溶血性弧菌的鉴定及同源性分析的性能.方法 将50株海洋副溶血性弧菌以及与其亲缘关系相近的30株海洋溶藻弧菌、8株海洋坎贝式弧菌、11株海洋哈维氏弧菌、3株海洋需钠弧菌、2株海洋创伤弧菌分别用16S rRNA、rpoB和MALDI-TOF MS技术3种方法进行鉴定并做系统进化树、聚类分析,比较分析结果.结果 50株海洋副溶血性弧菌用16S rRNA鉴定出5株,rpoB鉴定出41株,MALDI-TOF MS技术鉴定出41株.16S rRNA系统进化树的结果不能将海洋副溶血性弧菌和海洋溶藻弧菌、海洋坎贝式弧菌、海洋需钠弧菌、海洋创伤弧菌分开.rpoB和MALDI-TOF MS可以准确地做出同源性分析.rpoB鉴别来自不同地域的海洋副溶血性弧菌的能力优于MALDI-TOF MS,而MALDI-TOF MS在海洋副溶血性弧菌的环境株和临床株的鉴别上优于rpoB.结论 rpoB和MALDI-TOF MS技术鉴定海洋副溶血性弧菌准确,二者相结合是进行海洋副溶血性弧菌鉴定和同源性分析的适宜技术.%Objective To investigate the application of 16S ribosomal RNA gene (rRNA),RNA polymerase β-subunit gene(rpoB)and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) in identification and homology analysis of marine bacteria vibrio parahaemolyticus.Methods Fifty strains of marine vibrio parahaemolyticus and 54 strains of pathogens with close genetic relation (30 marine vibrio alginolyticus,8 marine vibrio campbellii,11 marine vibrio harveyi,3 marine vibrio natriegen and 2 marine vibrio vulnificus) were isolated and detected by 16S rRNA,rpoB and MALDI-TOF MS techniques.Results Five isolates of vibrio parahaemolyticus were identified by 16S rRNA technique;41 isolates were identified by rpoB and MALDI-TOF MS techniques,respectively.The phylogenetic tree constructed by 16S rRNA could not discriminate vibrio parahaemolyticus from vibrio alginolyticus,vibrio campbellii,vibrio natriegen and vibrio vulnificus;rpoB and MALDI-TOF MS could identify homological pathogens.The discrimination ability of rpoB was better than that of MALDI-TOF MS in identifying marine vibriones from different regions;MALDI-TOF MS was superior to rpoB in distinguishing environmemal and clinical vibrio parahaemolyticus strains.Conclusion RpoB and MALDI-TOF MS are accurate and appropriate techniques for identification and homology analysis of marine bacteria vibrio parahaemolyticus.
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