Description:1.IntendedUse TheGenWayBordetellapertussisIgGAntibodyELISATestKithasbeendesignedforthethe detectionandthequantitativedeterminationofspecificIgGantibodiesagainstBordetellapertussis inserumandplasma.FurtherapplicationsinotherbodyfluidsarepossIBLeandcanberequested fromtheTechnicalServiceofGenWay. Thisassayisintendedforresearchuseonly. 2.GeneralInformation WhoopingcoughisadiseaseoftherespiratorytractswhichiscausedbyBordetellapertussis bacteria.Itistransmittedbyairborneinfection.ThegramnegativeCoccobacillusproducesaseries ofBIOLOGicallyactivemolecules.Thedifferentcompoundsappeareitherduringthepathogenesisor duringtheprocessofimmunizationagainstpertussisandshowdifferenteffects.Acharacterisation hasbeenmadeforthepertussistoxin(pt),the filamenteryhaemagglutinine(fha)anddifferent lipopolysaccharides(lps). Pertussisshowsahighrateoftransmission(ratesofinfectionofover90%havebeenfoundfor non-vaccinatedhouseholdmembers)andcancauseseverediseases,especiallyforveryyoung children.From10749individualsunderoneyearbetween1980and198969%werebroughtinto hospital,22%sufferedfrompneumonia,0.9%showedanEncephalopathyand0.6%died. Forolderchildrenandadults(includingalreadyvaccinatedpersons)theinfectionmaybeobserved byanunspecifiedbronchitisorinflammationoftheupperrespiratorytracts.Evenasymptomatic casesarequitecommon. TheSEROlogicalresponsefollowingpertussisdiseaseorimmunizationwithpertussisvaccinehas beenmeasuredwithagglutinationassays,precipitins,complementfixationandenzyme-linked immunosorbentassay(ELISA). Enzyme-linkedimmunosorbentassays,inwhich Bordetellaantigen(containingtoxin,FHAand LPSandstandardizedinU/ml)isboundtoasolidphasesupport,aresensitive,easytoperformand canbeusedbothtodetermineseropositivitywithasingleserumandtoindicaterecentBordetella infectionbydeterminationofIgMandIgA. 3.PrincipleoftheTest TheGenWayBordetellapertussisIgGantibodytestkitisbasedontheprincipleoftheenzyme immunoassay(EIA).Bordetellaantigenisboundon thesurfaceofthemicrotiterstrips.Diluted sampleserumorready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.Abinding betweentheIgGantibodiesoftheserumandtheimmobilizedBordetellaantigentakesplace.After aonehourincubationatroomtemperature,theplateisrinsedwithdilutedwashsolution,inorderto removeunboundmaterial.Thenready-to-useanti-human-IgGperoxidaseconjugateisaddedand incubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedand incubatedfor20minutes,inducingthedevelopmentofabluedyeinthewells.Thecolor developmentisterminatedbytheadditionofastopsolution,whichchangesthecolorfromblueto yellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelengthof450nm.The concentrationoftheIgGantibodiesisdirectlyproportionaltotheintensityofthecolor. AdditionalInformation:NameBordetellapertussisIgGELISARelatedProductNamesBordetellapertussisIgGELISAApplicationsEnzymeimmunoassaybasedonmicrotiterplateforthedetectionandquantitativedeterminationofhumanIgGantibodiesagainstBordetellapertussisinserumandplasmaIntendedUseResearchUseOnly