Description:ProductName:RubellaIgMELISAIntendedUse:TheGenWayRubellaIgMAntibodyELISATestKithasbeendesignedforthedetectionandthequantitativedeterminationofspecificIgMantibodiesagainstRubellavirusinserumandplasma.FurtherapplicationsinotherbodyfluidsarepossIBLeandcanberequestedfromtheTechnicalServiceofGENWAY.Monitoringofpatientswithengraftedorgansaswellasdeterminationofantibodiesinotherbodyfluidswerenotvalidatedandhencearewithintheresponsibilityoftheuser.Thisassayisintendedforresearchuseonly.Laboratoryresultscanneverbetheonlybaseofamedicalreport.Thepatienthistoryandfurthertestshaveadditionallytobetakenintoaccount.GeneralInformation:Rubellainfectionbelongstotheclassicalchildren"sdiseaseswithalife-longimmunity,andthevirusisspreadworldwideendemically.Innon-vaccinatedpopulations,80-90%oftheinfectionsoccurduringthechildhood.Inspiteoftherubellavaccination,introducedin1974,inGermanytherecontinuetoappearconnataldiseases.ThecausativeagentisageneticallystableRNAvirus,whichbelongstothegenusrubiviruswithinthefamilyoftogaviridae.Humanbeingsaretheonlyknownnaturalhostsfortherubellavirus.Thetransmissionoccursviadropletinfection,withanincubationtimeof14-23days.Clinicallythediseasemanifestsitselflikealightfluinfection.Thenucalandretroauricallymphnodesareswollen,andamoderateenlargementofthespleenisobserved.Ashortandmediumraiseoftemperatureappearstogetherwitharatherslightsensationofillness.Rubellaisovercomeeasilywithinsignificantandlightsymptomsduringthechildhood,howevermoreattentionisrequiredinthecaseoftheinfectionofnon-immunizedpregnantwomen,becauseofthepossiblemalformationsofthefoetus,whichcanbegenerated.Astheinfectioncanbetransmittedviatheplacenta,thedevelopingfoetuscansufferseveredamages,thefrequencyandgravityofwhichisdependentonthemomentofinfectionduringpregnancy.Arubellainfectionduringthe1sttill4thmonthcanleadtoaspontaneousabortionorprematurebirth.Sinceaspecificcausaltherapydoesnotexist,thesecondarysignslikefever,arthritisorarthralgiesaretreatedsymptomatically.Theclinicaldifferentialdiagnosisisproblematic,becausesimilarexanthemsandfeverishillnessesappearalsointhecourseofotherchildren"sdiseaseslikemeasles,scarletandparvovirusitis.Thefollowinglaboratorymethodsareavailable:hemagglutinationinhibitiontest(HIT),hemolysisin-geltestorELISA.Thedetectionofvirus-specificIgMantibodiesisimportantfortheassessmentoffreshinfections,andtheIgGtestisusedforthedeterminationofimmunity.Inthecaseofsevereconnatalinfections,theisolationoftherubellavirusfrompharyngeallavage,urineandothersecretionscanalsobeperformed.PrincipleoftheTest:TheGENWAYRubellaIgMantibodytestkitisbasedontheprincipleoftheenzymeimmunoassay(EIA).Rubellavirusantigenisboundonthesurfaceofthemicrotiterstrips.Dilutedpatientserumorready-to-usecalibratorsarePipettedintothewellsofthemicrotiterplate.AbindingbetweentheIgMantibodiesoftheserumandtheimmobilizedRubellavirusantigentakesplace.Afteraonehourincubationatroomtemperature,theplateisrinsedwithdilutedwashsolution,inordertoremoveunboundmaterial.Thenready-to-useanti-human-IgMperoxidaseconjugateisaddedandincubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedandincubatedfor20minutes,inducingthedevelopmentofabluedyeinthewells.Thecolordevelopmentisterminatedbytheadditionofastopsolution,whichchangesthecolorfrombluetoyellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelengthof450nm.TheconcentrationoftheIgMantibodiesisdirectlyproportionaltotheintensityofthecolor.Storageandlimitationofusage:(expirydatesareprintedonthelabels)Storethecomponentsofthekitat2-8°C.Afterusageputtheplateintheplasticbag,closethebottleswiththeirscrewcaps,andagainstorethekitat2-8°C.Afterthefirstopeningthekitshouldbeusedwithin3months,thedilutedwashbuffercanbekeptfor4weeksat2-8°C.AdditionalInformation:NameRubellaIgMELISARelatedProductNamesRubellaIgMELISAIntendedUseResearchUseOnly