Description:1.IntendedUse TheGenWayTBEVIgGAntibodyELISATestKithasbeendesignedforthethedetectionandthe quantitativedeterminationofspecificIgGantibodiesagainstTBEVinserumandplasma.Further applicationsinotherbodyfluidsarepossIBLeandcanberequestedfromtheTechnicalServiceof GenWay. Thisassayisintendedforresearchuseonly. 2.GeneralInformation TheinfectionwithTBEV(tick-borneencephalitis virus)occurspredominantlyinearlysummer throughthebiteofatick,infectedwithTBEV.Tickscanbecomeinfectedduringanystageoftheir metamorphosisandtransmitthevirusdiaplacentally.Itisassumedthatthevirushibernatesinthe tickorothervertebrateslikeporcupines,dormice,batsanddeer.Theincubationtimerangesfrom4 -21days.Theviruseswereearliercalledarboviruses(arthropod-borneviruses)andarecombined bytheirfunctionintoagroupofinfectiouspathogens,beingtransmittedbetweenvertebratesbythe biteofblood-suckingarthropods.Thesevirusesreplicateintheorgansofthereceivingarthropods, withoutcausingsymptomsofadiseaseortissuedamage.Thecyclusvertebrate-arthropod-vertebrateiskeptalivebythisvectorwhichbecomeslife-longinfectiousthroughtheblood-suckingof avertebrateintheviraemicphase.Today,TBEVisallocatedtothegroupofflaviviruses. SEROlogically,TBEVbelongstoflavivirusgroupB.ThepathogensconsistofasinglestrandRNA andareinactivatedbyetherandsodiumdesoxycholate.Theviruscanbemultipliedinthebrainof newbornmice,butalsoincellcultures(BHK-21andVerocells).Theviralreplicationtakesplace inthecytoplasmaofinfectedcells.ThevirionsformintheGolgiregion.Theparticlesizeofthe virusesis80-100nm.ThevirusesaretransmittedbythetickIxodesricinus.Characteristicforthe diseaseisaninvolvementofthebulbaryorcervicalsectionsofthespinalmarrowaswellasthe developmentofascendingparalysisorhemiparesis.Theoccurenceofresidualparaplegiaafterthe infectionisnotuncommon.Typicalinfectionsshowatwo-stagecourseofthedisease:thefirst phasecomesalongwithanuncharacteristicfeverishinfectwithstrongheadaches, thesecondphase withameningo-encephalitiswithorwithoutpareses.Afteranincubationtimeof3-28days (dependingonthedosisoftheinfection),theinitialphasedevelopswithunspecificgrippal symptoms.3-7dayslater,thesymptomsdecreaseor,inapproximately10%ofthecases,thesecondstageofthediseasedevelops,markedbyarenewedincreaseoftemperature,headache,painin thelimbsandmeningo-encephaliticsymptoms.Paralysisinthescapularareamayoccur.Afew dayslater,astageofimprovementstartswhichgenerallylasts1-3weeks.IftheCNSisinvolved, residualdamageoccursinabout7%ofthecases,letalityisaround1%.Theclinicalpictureof TBEVisuncharacteristic,thus,aserologicallaboratorydiagnosisisrequired.Thediagnosisofa freshinfectionisdonebythedetectionofspecificIgMantibodies. TBEV-specificIgGantibodiesaredeterminedfortworeasons: a.)tomonitorthecourseofthediseaseafterinfectionwiththeTBEV(significanttiterchanges) b.)todeterminethe&BDquo;immunestatus“afteran„activeimmunization“(vaccinationprotection) TheenzymeimmunoassayforthedetectionofTBEVishighlysensitiveandallowsthe determinationofTBEV-specificantibodiesinsamplesera.3.PrincipleoftheTest TheGenWayTBEVIgGantibodytestkitisbasedontheprincipleoftheenzymeimmunoassay (EIA).TBEVantigenisboundonthesurfaceofthemicrotiterstrips.Dilutedsampleserumor ready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.AbindingbetweentheIgG antibodiesoftheserumandtheimmobilizedTBEVantigentakesplace.Afteraonehourincubation atroomtemperature,theplateisrinsedwithdilutedwashsolution,inordertoremoveunbound material.Thenready-to-useanti-human-IgGperoxidaseconjugateisaddedandincubatedfor30 minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedandincubatedfor20 minutes,inducingthedevelopmentofabluedyeinthewells.Thecolordevelopmentisterminated bytheadditionofastopsolution,whichchangesthecolorfrombluetoyellow.Theresultingdyeis measuredspectrophotometricallyatthewavelengthof450nm.TheconcentrationoftheIgG antibodiesisdirectlyproportionaltotheintensityofthecolor.AdditionalInformation:NameTBEV(FSME)IgGELISARelatedProductNamesTBEV(FSME)IgGELISAIntendedUseResearchUseOnly