应用预防医学2016年10月第22卷第5期 J Applied Prev Med. October2016.vd22Na5 论著· 文章编号:1673-758X201605-0386-04 中图分类号:R117 文献标识码:A 4株气单胞菌临床分离株基于gyB和rpoB基因序列的分子鉴定 韦小瑜',游旅',田克诚',朱琳,张仁俊3,吉光辉3,唐光鵬',王定明 1贵州省疾病预防控制中心传染病防治研究所(贵州贵阳50004) 2贵州省遵义市疾病预防控制中心(貴州遵义563000 [摘要]目的对4株气单胞菌临床分离株进行菌种的分子鉴定,为气单胞菌的种型鉴定提供病原学依据。方法对 4株系统生化鉴定为气单胞菌的菌株,提取核酸作为模板,分别使用srB、叩oB基因进行扩增及产物测序,将所得序 列在 GenBank中进行同源性搜索,与 Genbank中已经公布的同源性较高的气单胞菌序列比较构建系统发育树并进行分 析。结果4株气单胞菌gyTB基因序列经同源性搜索,与多株嗜水气单胞菌和豚鼠气单胞菌同源性为99%,moB基因 序列则与多株豚鼠气胞菌同源性为99%~100%,对rB基因和rpoB基因序列分别进行系统发育树分析显示,4株气 单胞菌菌株在grB基因系统发育树中与多株嗜水气单胞菌和豚鼠气单胞菌聚为一簇,而在oB基因系统发育树中与 多株豚鼠气单胞菌聚在同一分枝内。结论4株气单胞菌经mpoB基因测序分析明确鉴定为豚鼠气单胞菌,grB基因测 序分析不能有效鉴定其种型。 [关键词]气单胞菌;基因种;鉴定 Molecular identification of 4 clinical Aeromonas isolates by gyrB and rpoB gene sequencing WEI Xiao-yu, YOU Lv, and TIAN Ke-cheng, et al. Research Institute of Infectious Disease Control and Preventie Guizhou Provincial Center for Disease Control and Prevention, Guiyang 550004. China: Zuiyi Center for Disease Control and Prevention. Zunyi 563000. China [Abstract] Objective To identify the gene species of 4 clinical Aeromonas isolates and to provide the etiologic basis for the identification of Aeromonas species. Methods The isolates were identified as Aeromonas by conventional biochemical methods. The DNA of Aeromonas isolates was extracted as the template of PCR. The gyrB and rpoB gene were amplified and sequenced. Two sequence homology searches were conducted on Gen Bank and selected closely homology strains. Two molecular phylogenetic trees were constructed for analyze. Results The sequence homology of gyrB gene was 99%o among Aeromonas isolates, Aeromonas hydrophila and Aeromonas caviae strains, 99%100%(rpoB) among Aeromonas isolates and Aeromonas caviae strains. The phylogenetic tree of gyrB gene showed that 4 Aeromonas isolates, Aeromonas caviae strains and Aeromonas hydrophila strains wer clustered in the same group. While 4 Aeromonas isolates fell into the same branch with Aeromonas caviae strains the phylogenetic tree of rpoB gene. Conclusions Four Aeromonas isolates were identified as Aeromonas caviae by rpoB gene sequence. Aeromonas species could not be identified by gyrB gene sequence [Key Words] Aeromonas: genospecies; identification 基金项目:国家科技重大专项(2012ZX10004-212);贵州 气单胞菌属是一类革兰阴性杆菌,常引起鱼 省优秀青年科技人才培养对象专项资金项目黔科合人字 (2015)09号;贵州省疾病预防控制中心科技基金 类、鸟类及哺乳类等多种动物发病及感染人类引起 (2015-E2-3)联合资助 腹泻、败血症等,属典型的人兽共患病病原菌 作者简介:韦小瑜(1978—),女(苗族,贵州遵义人,副气单胞菌引起的败血症,主要发生于肝硬化及恶 主任医师,主要从事肠道传染病的实验室诊断和分子流行性肿瘤等免疫功能低下的患者,预后差,病死率 高。随着旅游业的发展和海产品食用量的增加应用预防医学 2016年10月第 22 卷第5期 J Applied Prev Med, October 2016, Vol 22 No.5 韦小瑜1 ，游旅1 ，田克诚 1 ，朱琳2 ，张仁俊2 ，吉光辉2 ，唐光鹏1 ，王定明1 1 贵州省疾病预防控制中心传染病防治研究所（贵州 贵阳 550004） 2 贵州省遵义市疾病预防控制中心（贵州 遵义 563000） 4株气单胞菌临床分离株基于gyrB和rpoB基因序列的分子鉴定 文章编号：1673 - 758X(2016)05 - 0386 - 04 中图分类号：R 117 文献标识码：A ·论著· [摘要] 目的 对4株气单胞菌临床分离株进行菌种的分子鉴定，为气单胞菌的种型鉴定提供病原学依据。方法 对 4株系统生化鉴定为气单胞菌的菌株，提取核酸作为模板，分别使用gyrB、rpoB基因进行扩增及产物测序，将所得序 列在GenBank中进行同源性搜索，与GenBank中已经公布的同源性较高的气单胞菌序列比较构建系统发育树并进行分 析。结果 4株气单胞菌gyrB基因序列经同源性搜索，与多株嗜水气单胞菌和豚鼠气单胞菌同源性为99%，rpoB基因 序列则与多株豚鼠气胞菌同源性为99%～100%，对gyrB基因和rpoB基因序列分别进行系统发育树分析显示，4株气 单胞菌菌株在gyrB基因系统发育树中与多株嗜水气单胞菌和豚鼠气单胞菌聚为一簇，而在rpoB基因系统发育树中与 多株豚鼠气单胞菌聚在同一分枝内。结论 4株气单胞菌经rpoB基因测序分析明确鉴定为豚鼠气单胞菌，gyrB基因测 序分析不能有效鉴定其种型。 [关键词] 气单胞菌；基因种；鉴定 基金项目：国家科技重大专项 （2012ZX10004-212）；贵州 省优秀青年科技人才培养对象专项资金项目[黔科合人字 （2015） 09 号]； 贵 州 省 疾 病 预 防 控 制 中 心 科 技 基 金 （2015-E2-3）联合资助。 作者简介：韦小瑜 （1978—），女(苗族)，贵州遵义人，副 主任医师，主要从事肠道传染病的实验室诊断和分子流行 病学研究。 Molecular identification of 4 clinical Aeromonas isolates by gyrB and rpoB gene sequencing WEI Xiao-yu, YOU Lv, and TIAN Ke-cheng, et al. Research Institute of Infectious Disease Control and Prevention, Guizhou Provincial Center for Disease Control and Prevention, Guiyang 550004, China; Zuiyi Center for Disease Control and Prevention, Zunyi 563000, China [Abstract] Objective To identify the gene species of 4 clinical Aeromonas isolates and to provide the etiologic basis for the identification of Aeromonas species. Methods The isolates were identified as Aeromonas by conventional biochemical methods. The DNA of Aeromonas isolates was extracted as the template of PCR. The gyrB and rpoB gene were amplified and sequenced. Two sequence homology searches were conducted on GenBank and selected closely homology strains. Two molecular phylogenetic trees were constructed for analyze. Results The sequence homology of gyrB gene was 99% among Aeromonas isolates、 Aeromonas hydrophila and Aeromonas caviae strains,99%～100%(rpoB) among Aeromonas isolates and Aeromonas caviae strains. The phylogenetic tree of gyrB gene showed that 4 Aeromonas isolates、Aeromonas caviae strains and Aeromonas hydrophila strains were clustered in the same group. While 4 Aeromonas isolates fell into the same branch with Aeromonas caviae strains in the phylogenetic tree of rpoB gene.Conclusions Four Aeromonas isolates were identified as Aeromonas caviae by rpoB gene sequence. Aeromonas species could not be identified by gyrB gene sequence [Key Words] Aeromonas; genospecies; identification 气单胞菌属是一类革兰阴性杆菌，常引起鱼 类、鸟类及哺乳类等多种动物发病及感染人类引起 腹泻、败血症等，属典型的人兽共患病病原菌[1] 。 气单胞菌引起的败血症，主要发生于肝硬化及恶 性肿瘤等免疫功能低下的患者，预后差，病死率 高。随着旅游业的发展和海产品食用量的增加， 386