16S核糖体RNA基因和RNA聚合酶β亚基基因及基质辅助激光解析／电离飞行时间质谱鉴定海洋副溶血性弧菌及其同源性分析的研究

Identification and homology analysis of marine bacteria vibrio parahaemolyticus with 16S ribosomal RNA gene, RNA polymerase ~subunit gene and matrix-assisted laser desorption/ionization time-of-flight massspectrometry

目的评价16S核糖体RNA基因（16SrRNA）、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-TOFMS）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16SrRNA、rpoB和MALDI-TOFMS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16SrRNA鉴定出5株，rpoB鉴定出41株，MALDI-TOFMS技术鉴定出41株。16SrRNA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．TOFMS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-TOFMS。而MALDI-TOFMS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．TOFMS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 parahaemo- lyticus from vibrio alginolyticus, vibrio campbellii, vibrio natriegen and vibrio vulnificus ; rpoB and MALDI-TOF MS could identify homologlcal 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 environ- mental 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.