Description:1.IntendedUse TheGenWayBorreliaburgdorferiIgMantibodyELISAkithasbeendesignedforthedetectionand thequantitativedeterminationofspecificIgMantibodiesagainstBorreliaburgdorferiinserumand plasma.FurtherapplicationsinotherbodyfluidsarepossIBLeandcanberequestedfromthe TechnicalServiceofGenWay. Thisassayisintendedforresearchuseonly. 2.GeneralInformation BorreliaburgdorferibelongstothefamilyofspiRochetes,ofwhichthreetypeshavebeenidentified tobehumanpathogenic:Borreliaburgdorferisensustricto,B.afzeliiandB.garinii.Theendemic areasofBorreliaareinCentralandEasternEurope,aswellasRussia,ChinaandJapan.Theillness istransferredviatickbites,inEuropemainlybyIxodesricinus.Intheendemiczones,likeSouthern GermanyandAustria,upto50%oftheticksareinfected. Intheclinicalcourse,afteranerythemamigrans,e.g.withneuroborreliosis,whichappearsatthe firststage,alsochronicalarthritis,encephalitis,meningitis,myositisandhepatitisareobserved. Treatmentisdoneviadifferentantibiotics,e.g.doxycyclin,amoxicillin,cefuroximandpenicillinG. AspecificimmunizationispossiblewithimmunoprophylaxiseitherbyarecombinantOspAorbya recombinantpolyvalentOspCvaccine. Thelaboratorydiagnosisisperformedbythedetectionofantibodiesinbloodandcerebrospinal fluid.Methodsemployedare:ELISA,immunofluorescence,hemagglutinationorWesternblot. Besideswholecellextracts,recentlythereareincreasinglyusedpurifiedorrecombinantsingle proteinsasantigens.Thisbringshowevergenerallyadecreaseinsensitivity.Itcouldbeshownthat betweenthevarioustestmethodsthereappearsignificantdifferencesintheinterpretation,sothat themostreliablemethodseemstobethefollow-upofthetiterdevelopment.Westernblotservesas aconfirmatorytest,becauseelectrophoreticallyseparatedsingleantigenscanbeevaluatedintheir reactionwithspecificserumantibodies. TheGenWayBorreliaburgdorferiIgMELISAtestkitcontainsbesidesawholecellantigenextract ofBorreliaburgdorferisensustricto,whichcross-reactswithBorreliaafzeliiandBorreliagarinii, anadditionofpureOspC,whichincreasesthespecificityandsensitivityoftheassay.Ifthetest showsonlyapositiveIgMresult(IgMnegative),theimmunityoftheindividualcanbeconcluded. Asmentionedabove,inthecaseofaclearlypositiveclinicalfinding,thereshouldbeperformeda confirmationofthetiterincreasewiththesameassay.WithapositiveIgMatthesametime,thereis astrongsUSPicionofanactivedisease. 3.PrincipleoftheTest TheGenWayBorreliaburgdorferiIgMantibodytestkitisbasedontheprincipleoftheenzyme immunoassay(EIA).Borreliaburgdorferiantigenisboundonthesurfaceofthemicrotiterstrips. Dilutedsampleserumorready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.A bindingbetweentheIgMantibodiesoftheserumandtheimmobilizedBorreliaburgdorferiantigen takesplace.Afteraonehourincubationatroomtemperature,theplateisrinsedwithdilutedwash solution,inordertoremoveunboundmaterial.Thenready-to-useanti-human-IgMperoxidase conjugateisaddedandincubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB) solutionispipettedandincubatedfor20minutes,inducingthedevelopmentofabluedyeinthe wells.Thecolordevelopmentisterminatedbytheadditionofastopsolution,whichchangesthe colorfrombluetoyellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelength of450nm.TheconcentrationofIgMantibodiesisdirectlyproportionaltotheintensityofthecolor. AdditionalInformation:NameBorreliaburgdorferiIgMELISARelatedProductNamesBorreliaburgdorferiIgMELISAApplicationsEnzymeimmunoassaybasedonmicrotiterplateforthedetectionandquantitativedeterminationofhumanIgMantibodiesagainstBorreliaburgdorferiinserumandplasmaIntendedUseResearchUseOnly