中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2015年
1期
21-25
,共5页
弧菌,副溶血性%血清分型%电泳,凝胶,脉冲场%聚合酶链反应%多位点测序分型
弧菌,副溶血性%血清分型%電泳,凝膠,脈遲場%聚閤酶鏈反應%多位點測序分型
호균,부용혈성%혈청분형%전영,응효,맥충장%취합매련반응%다위점측서분형
Vibrio parahaemolyticus%Serotyping%Electrophoresis,gel,pulsed-field%Polymerase chain reaction%Multi locus sequence typing
目的 分析2006-2012年深圳市食物中毒中分离的副溶血弧菌O3∶K6血清变异型毒力基因和分子分型特征,及其与O3∶K6血清型的基因遗传关系.方法 对深圳市2006-2012年食物中毒病例肛拭子样本中分离的1 15株副溶血弧菌进行PFGE分型,并根据地域分布与分离时间的不同,选取了7株O3∶K6血清型为对照组,对其以及O3∶K6血清变异型分别进行tdh和trh毒力基因检测,群特异性PCR (GS-PCR)和orf8基因检测,及多位点测序分型(MLST).结果 115株副溶血弧菌中,带型相似度＞80％的菌株占58.3％(67/115),其中,O3∶K6血清型占67％ (44/67),O1∶KUT血清型占5％(4/67),O3∶K6血清变异型占28％(19/67).O3∶K6血清变异型中,O1∶ K25血清型占7％(5/67),O4∶K68血清型占10％ (7/67),O11∶K36血清型占10％ (7/67),其均携带tdh基因,不携带trh基因,GS-PCR扩增基因和orf8基因阳性占10/19,GS-PCR扩增基因和orf8基因阴性占5/19,GS-PCR扩增基因阴性、orf8基因阳性占4/19,ST-3型占17/19,ST-305型占2/19.对照组均携带tdh基因,不携带trh基因,为GS-PCR扩增基因、orf8基因阳性以及ST-3型.ST-305型与ST-3型在dtdS和PntA两个管家基因上有差异,在dtdS基因的第305个碱基处,ST-305型(147号等位基因)为T,ST-3型(4号等位基因)为C;在pntA基因的第33个碱基处,ST-305型(93号等位基因)为C,ST-3型(29号等位基因)为A.结论 04∶K68、O1∶K25、O11∶K36血清型毒力基因携带与O3∶K6血清型相似,且MLST结果一致,遗传关系接近,为O3∶K6血清变异型.
目的 分析2006-2012年深圳市食物中毒中分離的副溶血弧菌O3∶K6血清變異型毒力基因和分子分型特徵,及其與O3∶K6血清型的基因遺傳關繫.方法 對深圳市2006-2012年食物中毒病例肛拭子樣本中分離的1 15株副溶血弧菌進行PFGE分型,併根據地域分佈與分離時間的不同,選取瞭7株O3∶K6血清型為對照組,對其以及O3∶K6血清變異型分彆進行tdh和trh毒力基因檢測,群特異性PCR (GS-PCR)和orf8基因檢測,及多位點測序分型(MLST).結果 115株副溶血弧菌中,帶型相似度＞80％的菌株佔58.3％(67/115),其中,O3∶K6血清型佔67％ (44/67),O1∶KUT血清型佔5％(4/67),O3∶K6血清變異型佔28％(19/67).O3∶K6血清變異型中,O1∶ K25血清型佔7％(5/67),O4∶K68血清型佔10％ (7/67),O11∶K36血清型佔10％ (7/67),其均攜帶tdh基因,不攜帶trh基因,GS-PCR擴增基因和orf8基因暘性佔10/19,GS-PCR擴增基因和orf8基因陰性佔5/19,GS-PCR擴增基因陰性、orf8基因暘性佔4/19,ST-3型佔17/19,ST-305型佔2/19.對照組均攜帶tdh基因,不攜帶trh基因,為GS-PCR擴增基因、orf8基因暘性以及ST-3型.ST-305型與ST-3型在dtdS和PntA兩箇管傢基因上有差異,在dtdS基因的第305箇堿基處,ST-305型(147號等位基因)為T,ST-3型(4號等位基因)為C;在pntA基因的第33箇堿基處,ST-305型(93號等位基因)為C,ST-3型(29號等位基因)為A.結論 04∶K68、O1∶K25、O11∶K36血清型毒力基因攜帶與O3∶K6血清型相似,且MLST結果一緻,遺傳關繫接近,為O3∶K6血清變異型.
목적 분석2006-2012년심수시식물중독중분리적부용혈호균O3∶K6혈청변이형독력기인화분자분형특정,급기여O3∶K6혈청형적기인유전관계.방법 대심수시2006-2012년식물중독병례항식자양본중분리적1 15주부용혈호균진행PFGE분형,병근거지역분포여분리시간적불동,선취료7주O3∶K6혈청형위대조조,대기이급O3∶K6혈청변이형분별진행tdh화trh독력기인검측,군특이성PCR (GS-PCR)화orf8기인검측,급다위점측서분형(MLST).결과 115주부용혈호균중,대형상사도＞80％적균주점58.3％(67/115),기중,O3∶K6혈청형점67％ (44/67),O1∶KUT혈청형점5％(4/67),O3∶K6혈청변이형점28％(19/67).O3∶K6혈청변이형중,O1∶ K25혈청형점7％(5/67),O4∶K68혈청형점10％ (7/67),O11∶K36혈청형점10％ (7/67),기균휴대tdh기인,불휴대trh기인,GS-PCR확증기인화orf8기인양성점10/19,GS-PCR확증기인화orf8기인음성점5/19,GS-PCR확증기인음성、orf8기인양성점4/19,ST-3형점17/19,ST-305형점2/19.대조조균휴대tdh기인,불휴대trh기인,위GS-PCR확증기인、orf8기인양성이급ST-3형.ST-305형여ST-3형재dtdS화PntA량개관가기인상유차이,재dtdS기인적제305개감기처,ST-305형(147호등위기인)위T,ST-3형(4호등위기인)위C;재pntA기인적제33개감기처,ST-305형(93호등위기인)위C,ST-3형(29호등위기인)위A.결론 04∶K68、O1∶K25、O11∶K36혈청형독력기인휴대여O3∶K6혈청형상사,차MLST결과일치,유전관계접근,위O3∶K6혈청변이형.
Objective To characterize the O3∶K6 serovariant of Vibrio parahaemolyticus on virulence gene and molecular typing,and analyze the genetic relationship between O3∶K6 and O3∶K6 serovariants.Methods PFGE was performed on 115 strains of V.parahaemolyticus which were collected from the anal swab of cases of foodbrone diseases in Shenzhen during 2006-2012.According to isolation times and locations,7 strains of O3∶K6 were selected as control strains.Tdh gene,trh gene,orf8 gene were detected,GS-PCR,multi-locus sequence typing (MLST) were used to chracterize 7 strains of O3∶K6 and O3∶K6 serovariants.Results PFGE indicated that 58.3％ (67/115) of V.parahaemolyticus strains shared a high similarity of band pattern(similarity ＞ 80％),which comprised of O3∶K6 (44/67),O1∶ KUT(4/ 67),and O3∶ K6 serovariants (19/67).Among the O3∶ K6 serovariants,O1 ∶ K25 accounted for 7 ％ (5/67),O4∶K68 accounted for 10％ (7/67),Ol1∶K36 accounted for 10％ (7/67).They all carried both tdh and trh gene,and 53％ (10/19) was GS-PCR positive and carried orf8 gene,26％ (5/19) was both GS-PCR and orf8 gene negative,21％ (4/19) was GS-PCR negative,or.f8 gene positive,89％ (17/19) was assigned to ST-3,11％ (2/19) was assigned to ST-305.Seven strains of O3∶K6 was GS-PCR positive,carried off8 gene,assigned to ST-3.ST-305 and ST-3 had differences in 2 housekeeping genes,which was dtdS gene and pntA gene.In the 305th base of dtdS gene,ST-305(147 allele profile) was T,while ST-3(4 allele profile)was C.In the 33th base ofpntA gene,ST-305(93 allele profile) was T,while ST-3(29 allele profile) was C.Conclusion O4∶ K68,O1∶K25 and O11∶K36 were highly similar in virulencee gene carriage,MLST type of O3∶ K6,and aslo shared a close genetic relationship with O3∶ K6,thus were considered as O3∶K6 serovirants.
