CAJ | 학술논문

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.
목적 대량기식물중독조사중분리도적시만균속(Shewanella spp.)균주진행생화화분자생물학감정.방법 대2007년9월29일지10월3일간,마안산시적량기식물중독환자적항식、종업인원수식화잉여식물표본진행채집,안조국표방법(GB/T4789),대소유표본진행증균화선택성배양기분리배양,의사균락용VITEK-32화AP120E계통감정,용보조생화、생장、용혈화약민실험분석균주특성,동시확증16S rDNA병측서,용MEGA 4.0연건건립진화수병진행분군.결과 소유표본경증균후접충선택성배양기,공유8빈표본재TCBS화BP배양기상장출단일적균락,재삼당철경지(TSI)사면상적주요생장특정위:산류화경、불산기,양화매양성.8주균경VITEK-32감정의감정위해조시만균(S.algae)혹부패시만균(S.putrefaciens),경AP120E계통감정위부패시만균.재WS、SS화EMB배양기균미검측도시만균생장.비교시만균적16S rDNA서렬표명,기중7주위해조시만균,1주위부패시만균.몰유종저8빈표본중검측도기타장도병원균,포괄곽란호균、사문균、부용혈성호균、변형간균화금황색포도구균.결론 종식물중독환자중분리도시만균,위해균작위가능적식물중독병원균제공료선색.
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