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ConventionalPhenotypicConventionalPhenotypicAntibiogram抗菌Biotype生化分Phage 噬菌体分 分Minimalspanningtree(MSTree)ofMLSTdataon4257subspeciesEachcirclecorrespondstooneof1,095STs,whosesizeisproportionaltothenumberofMolecularfortrackingofVeterinaryVeterinaryPublicExperimentalanddemosessionsSamplepreparationsTrackingofmicrobialsources分子溯源X-scopescouldnot X-scopescouldnot iftheyareclonally??/Tracking/TrackingSystemTypeabilityDiscriminationReproducibilityofepidemicclones(确定流行株的 Totesthypothesesaboutthesourcesandvehiclesoftransmissionoftheseclones(流行株的传染源 Tomonitorthereservoirsoftheepidemic(病原是否还有其它中间宿主Post-epidemicsurveillance(流行后的监测Evaluationofcontrolmeasuresby eradication,ordecreasedcirculation,oftheepidemicclones(评价现有控制方法的成效)theprocessinglinesMolecularPFGE/foodborneIllnessoutbreaksrDNAgenerestriction s yPulsefieldgelelectrophoresis(PFGE,goldstandardinoutbreakinvestigation)Nucleicacidsequencing:intergenicorNucleicacidhybridizationofentireBasicpremiseisthatisolatesthatarepartofthesamechainoftransmissionareclonallyrelated–theprogenyofthesameancestorcell(基本假设:同一个细胞的子代在Extensivegenomicandphenotypicdiversityexistswithinmicrobialpopulationsofthesamespecies(同一种生物体的不同在组水平和表型上具有多样性)Clonallyrelatedisolatesexhibitsignificantlymoresimilarcharactersthanunrelatedstrains–epidemiologicalmarkers(具有”克隆”相关性的分离株之间比非相关Tystemsredesigndooptimiednnbtweenrltdndunrltdisoltsofthemirobeofintrst分型系统需要优化,以便能干区分目标病原在克隆上的相关与非相关株)MicrobialMicrobial:WhatdoesitAmatchmeansthecasesareMoreLikelytohaveacommonLessLikelytohaveacommonsourcethaniftheydidmatchDiscriminationIndex(分辨力N-thetotalnumberofs-thetotalnumberofnj-thenumberofisolatesbelongingtothejthGeneralprinciplesofmolecularsubtyusingrestriction..C......DNAsequencerecognizedbyrestrictionC=Restrictionendonuclease(“molecularCriteriaCriteriaforEvaluatingAschemeshouldideallybeversatilei.e.ofthemethodsystemshouldachieveaDIofofisolatesDiscriminationIndex(分辨力DIdependsonthenumberoftypesandonthehomogeneityofthefrequencydistributionofisolatesintotypes;Ideally,eachisolateshouldhaveadifferentForthepurposeofcalculation,nontypeableisolatesweregroupedPFGE:PFGE:s(PulseFieldGelDNAfingerprintsofDNAfingerprintsofproductionlineindistinguishableMeatProductionysisprocess:ListeriamonocytogenesPulsePulsefieldgel---Workflow&Time2StreakGrowatPourintotheMixwithagarosemould&solidify2-RestrictionenzymesLysewithdetergents&Plugswithtodigest treatwithproteinaseK24-36PFGEdataPFGEdataWhathappensWhathappenswithPulseNet:NextIncreasethenumberofPulseNetAhierltimeubty ndrltimenReducethetimeittakesforisolatestogofromclinicallabtoState/localpublichealthlabReducethetimeforPFGEtestingofCriticalfortimelydetectionofIncreasethelevelofcommunicationbetweenlaboratoriansandTimelyassignmentofPulseNetdesignationsforPFGEpatternsImprovebandmarkingamongallStrengthencollaborationswiththefoodFutureVibrioparahaemolyticus/V.YersiniaNew methodologyMultiple-locussequence PulseNetdatabasePulseNetPulseNetismovingthenucleotidelevelRawRaw特 的等 特定菌株等 ”集 StrainBFingerprintsEmergingtechnologyformolecularsubty :MLSTPFGE:PFGE:Advantages&MLSTMLSTforoutbreak?3PCRamplify7ofThegenefragmentssequenced(PCRproducts)AssigndifferentNos.fordiffernetalleles:1,2,3…Giveanallelictypeforall7locihavingthesameallelicComparisonofsequencetypes(STs)withthosein L.monocytogenesL.monocytogenesComparingPFGEwithDendrogramofDifferentStrainStrainStrainStrnStraiStrainStrainStrainPubMLSTPubMLSTPathogenPathogenTrackerLocationofLocationofgenesinthegenomesforMLSTVibriogenesforrecA(RecAdnaE(DNApolymeraseIII,alpha ChromosomeII:dtdS(ThreoninepyrCtnaANeighborjoiningNeighborUPMGAUPMGAumNeighborMinimumTheThekeyissueforWhichgenefragmentstoHouse- vsvirulenceWithinORFsorintergenicHowmanyfragmentsarerequiredtoprovideconvincingevidenceforsourcetrackingorepidemiologicalinvestigation?NeedmoregenomesequencingdataforagivenCamyplobacterCamyplobacterjejuni/coligenesfor
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