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1、EntericbacterianDemonstrationEntericbacteria(E.coli,Shigellae, Salmonellae, Proteus)nMorphology&stainingnGrowthcharacteristicsonSSSS agar plate and EMBagar plate and EMB agar agar plate)plate)nBiochemical reaction: kligerBiochemical reaction: kligers iron slant, s iron slant, Urease testUrease testn
2、OperationWidaltestLaboratorydiagnosisofentericbacteriamannitolfermentationtest（Continued）AntimicrobialSusceptibilitytest（Continued）n VideoAnaerobicbacteriaClassificationEnterobacteriaceaeShigellaEscherichiaSalmonellaProteusKlebsiellaETEC, EIEC, EPEC, EHEC, EAECS. dysenteriae,S.flexneri,S. boydii,S.
3、sonneiS. typhi, S.entericaBasiccharacteristicsnGram-negative rodnEasy to culture, form circular,convex, smooth colonies with distinct edges on basic medianBiochemical reaction: activenComplx antigenic structurenResistance: not strongnEasy to mutate Widaltestbeusedtohelpmakeapresumptivediagnosisofent
4、ericfevernPrinciple：S.typhi OantigenS.typhiHantigenS.paratyphiAHantigenS.schottmuelleriHantigenKnown antigensUnknown antibody in patient serumagglutination12345678S.typhi O0.2ml0.2mlS.typhi H 0.2ml0.2mlS.paratyphi A H0.2ml0.2mlS.Schottmuelleri H0.2ml0.2mlInitial dilution1:201:401:801:1601:3201:6401:
5、1280ControlSalineFinal dilution1:401:801:1601:3201:6401:12801:2560 Ag test Abknown unknownSerum 1:10 1:20 0.2ml4 0.8 ml 1.6mlSaline 0.8 ml mixing 0.8ml mixing 1:40 0.2ml4 Saline 0.8 ml 1.6ml 0.8ml mixing Saline 0.8 ml 1：80 Adding antigen1. Diluting Antibody(serum)2. Adding antigen3. Incubating at 37
6、, 60(at 45, 30)4. Reading the resultsLaboratory diagnosis of pathogenic Enteric bacteria (stool)2 suspected colonies2 suspected colonies(Small,colorless,transparent)InoculatedinKliglersironslantstoolSpecimen:SS agar platedifferential and selective medium Kliglers iron slantBiochemical reactionSlide
7、agglutinationserologictestCultivatedSS agar plate(Shigella & Salmonella) adifferentialandselectivemediumusedforinitialisolationofentericbacteriaContains：basicmedium,lactose,sodiumcitrate,sodiumthiosulfate,ironcitrate,bilesalts,brilliantgreen,neutralred(pH6.8-8.0, redpH6.8-8.0, redyellowyellow）Usage：
8、lactose,neutralred：nLactose fermented:fermentlactosewithsufficientacidproductiontochangetheneutralredformredcoloniesnLactose not fermented: colorless coloniesSodiumcitrate,brilliantgreen,bilesalts:inhibitE.coliResult:Result:E.coliShigellae & SalmonellaeRed,large,opaquecoloniesSmall,colorless,transpa
9、rentcoloniesEMB agar plate Contains：basicmedium,lactose,eosin,methyleneblueUsage：Lactose fermented:fermentlactosetoproduceacid,thenmakeeosinandmethylenebluetobeaBLACKcompound.Lactose not fermented: colorless coloniesResult:Result:E.coliShigellae & Salmonellaelarge,black，opaquecoloniesSmall,colorless
10、,transparentcolonies双糖管n接种方法n观察上层含乳糖部分n发酵乳糖致病菌红色，非致病菌黄色；nH2S试验大肠杆菌无黑色肖氏沙门菌出现黑色然后观察下层n发酵葡萄糖（产酸产气）及动力乳糖乳糖,Fe2+固体固体葡萄糖葡萄糖半固体半固体指示剂指示剂:酚红（酚红（pH6.8-8.4, 黄黄 红）红）Biochemicalreaction（Kliglersironslant改良克氏双糖铁琼脂斜面）nOberservationLactose（10）Glucose（1）Ferrous sulfateindicator:phenolredpH6.8-8.4pH6.8-8.4YellowYel
11、lowredredGlucoselactoseH H2 2S S+-+SlantYellowRedblackBottomYellowandbubbleYellowblackInocuatingKliglersironagarslantnInoculatingneedlenRemovingcellsfromssagarplatewithaneedlenStabneedletobottomofthetubenAstheneedleiswithdrawnfromtheagar,streakitacrossthefaceoftheslantfrombottomtothetopnIncubateat37
12、 Lactose Glucose H2S Urease E.coli +S. dysenterias + -S. typhi + +/-S. paratyphi A + -Proteus + + Laboratory diagnosis of pathogenic Enteric bacteria (stool)stoolSpecimen:SS agar platedifferential and selective medium Kliglers iron slantBiochemical reactionSlide agglutinationserologictestReadingther
13、esultsofWidaltestnExaminingthecontroltubesfirst:negativenTheagglutinationofOantigenasa“matt”or“carpet”atthebottom.nTheagglutinationofHantigensloose,woolyorcottonyn（）（）liquid clear, all antigen are agglutinate at the bottom.（）（） liquid clear, most of antigen(75%) are agglutinate at the bottom（）（） liq
14、uid semitransparent , part of antigen are agglutinate at the bottom obviously.（）（） liquid turbid, a little antigen are agglutinate at the bottom.（）（）negative, same as controlAgglutination titer: the highest dilution of “+”Interpretationofwidaltestnbaseline：S.typhi Oantibody 1：80， S.typhi Hantibody 1：160, S.paratyphiAHantibody 1：80nA rise in titre between two sera specimens is more meaningful than a single test.0 antibodyH antibodyInterpretationHighHighentericfeverNormalNormalNormalNormalHighsufferedinthepast,vaccinatedHighNormalintheearlystages,Cross-reactionTheEndThanks
