Description:Enzymeimmunoassaybasedonmicrotiterplate forthedetectionandquantitativedetermination ofhumanIgMantibodiesagainstYersiniaenterocoliticainserumandplasma  IntendedUse TheGenWayYersiniaIgMAntibodyELISATestKithasbeendesignedforthethedetectionand thequantitativedeterminationofspecificIgMantibodiesagainstYersiniaenterocoliticainserum andplasma.FurtherapplicationsinotherbodyfluidsarepossIBLeandcanberequestedfromthe TechnicalServiceofGenWay. Thisassayisintendedforresearchuseonly.GeneralInformation ThepathogenicgermsYersiniapestis,Y.pseudotuberculosis,Y.enterocoliticaandY.ruckeri belongtothegenusYersiniaasamemberof theenterobacteriaceafamily.Allthespeciesof medicalimportanceoccurfacultativelyinsidethecells,whichleadstothecharacteristical inflammationofthelymphatictissueinthecourseofanillness.Yersiniaenterocoliticaistakenup orally,andthesymptomsinanindividualareterminalileitisaswellasdiarrhoea.Itisdifficultto makeaseparationfromappendicitisbydifferentialdiagnosis.Inthecourseofaretarded immunologicalreaction,extraintestinalmanifestationslikeerythemanodosum,uveitis,andarthritis canappear.IthasbeenclaimedthatthebackgroundforareactivearthritiscausedbyYersinia consistsinthelocalsynthesisofantibodiesinthejoints(synovialfluid). Yersiniacanbecharacterizedwithoutproblemsbystandardizedbacterialtests,afterisolationina pureculture.Followingahumandisease,abovealltheSEROtypesO3,O8andO9arefound.CrossreactivitiestoBrucellaaredescribedandhavetobetakenintoaccountbythedifferentialdiagnosis. TheclassicalserologicaldetectionmethodhasalwaysbeentheWidaltest.Furtherincreasingly popularmethodsaretheHAT,theKBRtestaswellastheELISA.Theenzymeimmunoassayis characterizedontheonehandsidebyahighsensitivity,butalsobythepossibilitytodifferentiate betweenIgGandIgA/IgMantibodies.TheimmunoglobulinclassesIgAandIgMshouldbereferred toasacriteriumofinterpretationforanactiveprocess,whenanarthriticdiseaseissUSPected. Becausethereexistsalsoanimmunologicalsimilaritywiththethyrotropinreceptor,thetestcan alsobeemployedfortheconfirmationoftheautoimmuneGraves’disease.Asaconfirmatorytest withthepossibilityofanidentificationofseparatebacterialproteinsaswellasfortheexclusionof cross-reactivities,theWesternBlotmethodhasprovedsuccessful.PrincipleoftheTest TheGenWayYersiniaenterocoliticaIgMantibodytestkitisbasedontheprincipleoftheenzyme immunoassay(EIA).Yersiniaantigenisboundon thesurfaceofthemicrotiterstrips.Diluted sampleserumorready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.Abinding betweentheIgMantibodiesoftheserumandtheimmobilizedYersiniaantigentakesplace.Aftera onehourincubationatroomtemperature,theplateisrinsedwithdilutedwashsolution,inorderto removeunboundmaterial.Thenready-to-useanti-human-IgMperoxidaseconjugateisaddedand incubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedand incubatedfor20minutes,inducingthedevelopmentofabluedyeinthewells.Thecolor developmentisterminatedbytheadditionofastopsolution,whichchangesthecolorfromblueto yellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelengthof450nm.The concentrationoftheIgMantibodiesisdirectlyproportionaltotheintensityofthecolor. AdditionalInformation:NameYersiniaIgMELISARelatedProductNamesYersiniaIgMELISAIntendedUseResearchUseOnly