Description:EnzymeimmunoassaybasedonmicrotiterplateforthedetectionandquantitativedeterminationofhumanIgAantibodiesagainstYersiniaenterocoliticainserumandplasmaIntendedUseTheGenWayYersiniaIgAAntibodyELISATestKithasbeendesignedforthethedetectionandthequantitativedeterminationofspecificIgAantibodiesagainstYersiniaenterocoliticainserumandplasma.FurtherapplicationsinotherbodyfluidsarepossIBLeandcanberequestedfromtheTechnicalServiceofGenWay.Thisassayisintendedforresearchuseonly.GeneralInformationThepathogenicgermsYersiniapestis,Y.pseudotuberculosis,Y.enterocoliticaandY.ruckeribelongtothegenusYersiniaasamemberoftheenterobacteriaceafamily.Allthespeciesofmedicalimportanceoccurfacultativelyinsidethecells,whichleadstothecharacteristicalinflammationofthelymphatictissueinthecourseofanillness.Yersiniaenterocoliticaistakenuporally,andthesymptomsinanindividualareterminalileitisaswellasdiarrhoea.Itisdifficulttomakeaseparationfromappendicitisbydifferentialdiagnosis.Inthecourseofaretardedimmunologicalreaction,extraintestinalmanifestationslikeerythemanodosum,uveitis,andarthritiscanappear.IthasbeenclaimedthatthebackgroundforareactivearthritiscausedbyYersiniaconsistsinthelocalsynthesisofantibodiesinthejoints(synovialfluid).Yersiniacanbecharacterizedwithoutproblemsbystandardizedbacterialtests,afterisolationinapureculture.Followingahumandisease,abovealltheSEROtypesO3,O8andO9arefound.CrossreactivitiestoBrucellaaredescribedandhavetobetakenintoaccountbythedifferentialdiagnosis.TheclassicalserologicaldetectionmethodhasalwaysbeentheWidaltest.FurtherincreasinglypopularmethodsaretheHAT,theKBRtestaswellastheELISA.Theenzymeimmunoassayischaracterizedontheonehandsidebyahighsensitivity,butalsobythepossibilitytodifferentiatebetweenIgGandIgA/IgMantibodies.TheimmunoglobulinclassesIgAandIgMshouldbereferredtoasacriteriumofinterpretationforanactiveprocess,whenanarthriticdiseaseissUSPected.Becausethereexistsalsoanimmunologicalsimilaritywiththethyrotropinreceptor,thetestcanalsobeemployedfortheconfirmationoftheautoimmuneGraves’disease.Asaconfirmatorytestwiththepossibilityofanidentificationofseparatebacterialproteinsaswellasfortheexclusionofcross-reactivities,theWesternBlotmethodhasprovedsuccessful.PrincipleoftheTestTheGenWayYersiniaenterocoliticaIgAantibodytestkitisbasedontheprincipleoftheenzymeimmunoassay(EIA).Yersiniaantigenisboundonthesurfaceofthemicrotiterstrips.Dilutedsampleserumorready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.AbindingbetweentheIgAantibodiesoftheserumandtheimmobilizedYersiniaantigentakesplace.Afteraonehourincubationatroomtemperature,theplateisrinsedwithdilutedwashsolution,inordertoremoveunboundmaterial.Thenready-to-useanti-human-IgAperoxidaseconjugateisaddedandincubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedandincubatedfor20minutes,inducingthedevelopmentofabluedyeinthewells.Thecolordevelopmentisterminatedbytheadditionofastopsolution,whichchangesthecolorfrombluetoyellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelengthof450nm.TheconcentrationoftheIgAantibodiesisdirectlyproportionaltotheintensityofthecolor.References1.Bottone,E.J.(ed.).Yersiniaenterocolitica.CRCPress,BocaRaton(1981).2.Brubaker,R.R.ThegenusYersinia:biochemistryandgenetics.Curr.Top.Microbiol.,57:111(1972).3.Butler,T.PlagueandotherYersiniainfections.PlenumPublishingCorp.,NewYork(1983).4.Heesemann,J.etal.ExperimentalYersiniaenterocoliticainfections;amodelforhumanyersiniosis.APMIS,101:417(1993).5.Kittelberger,R.etal.Serologicalcross-reactivitybetweenBrucellaabortusandYersiniaenterocolitica.Vet.Microbiol.,47:271(1995).6.Luo,G.etal.PurificationandcharacterizationofYersiniaenterocoliticaproteins.J.Immunol.,152:2555(1994).7.Möaki-Ikola,O.etal.Yersinia-specificantibodiesinserumandsynovialfluid.Ann.Rheum.Dis.,53:535(1994).AdditionalInformation:NameYersiniaIgAELISARelatedProductNamesYersiniaIgAELISAIntendedUseResearchUseOnly