Abstract
汪永禄,王多春,詹圣伟,郑锦绣,刘燕,陶勇,石志峰,郝民,于礼,阚飙.从食物中毒患者标本中分离鉴定海藻和腐败施万菌[J].Chinese journal of Epidemiology,2009,30(8):836-840
从食物中毒患者标本中分离鉴定海藻和腐败施万菌
Isolation and characterization of Shewanella spp.from patients of food poisoning
Received:May 05, 2009  
DOI:
KeyWord: 食物中毒  施万菌属  分离  鉴定
English Key Word: Food poisoning  Shewanella spp.  Isolate  Characterization
FundProject:国家"863"高技术研究发展计划(2006AA02Z425);国家自然科学基金(30872260);国家"十一五"科技鼋大专项(2008ZX10004-012)
Author NameAffiliationE-mail
WANG Yong-lu Ma'anshan Center for Disease Control and Prevention, Ma'anshan 243000, China  
WANG Duo-chun 中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室  
ZHAN Sheng-wei Ma'anshan Center for Disease Control and Prevention, Ma'anshan 243000, China zhanshengwei@sina.com.cn 
ZHENG Jin-xiu Ma'anshan Center for Disease Control and Prevention, Ma'anshan 243000, China  
LIU Yan Ma'anshan Center for Disease Control and Prevention, Ma'anshan 243000, China  
TAO Yong Ma'anshan Center for Disease Control and Prevention, Ma'anshan 243000, China  
SHI Zhi-feng Ma'anshan Center for Disease Control and Prevention, Ma'anshan 243000, China  
HAO Min 中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室  
YU Li 中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室  
KAN Biao 中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室  
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Abstract:
      目的 对两起食物中毒调查中分离到的施万菌属(Shewanella spp.)菌株进行生化和分子生物学鉴定.方法 对2007年9月29日至10月3日间,马鞍山市的两起食物中毒患者的肛拭、从业人员手拭和剩余食物标本进行采集,按照国标方法(GB/T4789),对所有标本进行增菌和选择性培养基分离培养,疑似菌落用VITEK-32和AP120E系统鉴定,用辅助生化、生长、溶血和药敏实验分析菌株特性,同时扩增16S rDNA并测序,用MEGA 4.0软件建立进化树并进行分群.结果 所有标本经增菌后接种选择性培养基,共有8份标本在TCBS和BP培养基上长出单一的菌落,在三糖铁琼脂(TSI)斜面上的主要生长特征为:产硫化氢、不产气,氧化酶阳性.8株菌经VITEK-32鉴定仪鉴定为海藻施万菌(S.algae)或腐败施万菌(S.putrefaciens),经AP120E系统鉴定为腐败施万菌.在WS、SS和EMB培养基均未检测到施万菌生长.比较施万菌的16S rDNA序列表明,其中7株为海藻施万菌,1株为腐败施万菌.没有从这8份标本中检测到其他肠道病原菌,包括霍乱弧菌、沙门菌、副溶血性弧菌、变形杆菌和金黄色葡萄球菌.结论 从食物中毒患者中分离到施万菌,为该菌作为可能的食物中毒病原菌提供了线索.
English Abstract:
      Objective To identify the isolates of Shewanella spp.from specimens of food poisoning based on biological and biochemical analysis.Methods Strains were obtained from the investigation on two food poisoning episodes in September and October,2007 in Ma'anshan city,Anhui province.In accordance with the national standard protocol(GB/T 4789),all specimens were enriched and isolated on selective medium,and the suspected strains were ldentified by the VITEK-32 and API20E systems.For Shewanella spp.identified by the biochemical system,more characteristics were analyzed using auxiliary biochemical,growth,hemolytic and drug-resistance tests.DNAs of Shewanella spp.were extracted,16S rDNA was PCR amplified and sequenced with universal 16S rDNA primers.Phylogenetic tree was constructed with MEGA 4.0.Results After enrichment, all specimens were inoculated to selective medium and Shewanella spp.strains were isolated from 8 samples with single colony on both TCBS and BP media.The characteristics of growth in the Triple Sugar Iron (TSI) agar appeared to have had hydrogen sulfide production but no gas production or positive oxidase.No Shewanella spp.strain was detected in WS,SS and EMB media.The 8 strains were identified as Shewanella algae(S.algae) or Shewanella putrefaciens(S.putrefaciens) by VITEK-32,as S.putrefaciens by API20E system.No other enteropathogenic bacteria,including Vibrio cholerae,Salmonella,Vibrio parahaemolyticus,Proteus vulgaris or Staphylococcus aureua,were detected from those 8 samples.From 16S rDNA phylogenetic trees,7 out of 8 ShewaneUa spp.were identified as S.algae.1 as S.putrefaeiens.Conclusion Strains of Shewanella spp.were lsolated from samples of the food poisoning episodes,providing a possible clue to investigate the role of Shewanella spp.on food poisoning
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