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1.IntendedUse TheGenWayToxoplasmaIgGAntibodyELISATestKithasbeendesignedforthedetectionand thequantitativedeterminationofspecificIgGantibodiesagainstToxoplasmainserumandplasma. FurtherapplicationsinotherbodyfluidsarepossIBLeandcanberequestedfromtheTechnical ServiceofGenWay. Thisassayisintendedforresearchuseonly.
2.GeneralInformation Toxoplasmosisisaninfection,whichiscausedbytheparasitetoxoplasmagondii.Upto20%ofthe populationcarrythepathogenicagent,butonlyafewfeelsymptoms,becausetheimmunesystem hinderstheillnesstobreakout.Pregnantwomenshouldhoweverbeverycautious,asaninfection canbedetrimentaltothefoetus. Toxoplasmagondiibelongstotheprotozoaeandisaparasite,whichcaninfectmanydifferent speciesofmammals,amongstothershumanbeings.Animalslikecats,pigsandsheepspread oocystsaswellastissuecysts,whichafteringestionbyhumansareconvertedintotachyzoites, andthesecanbefoundafterwardsinnerveandmuscletissue. Whenapregnantwomanbecomes infected,thetachyzoitescanreachthefoetusviatheplacenta. Thesymptomsoftoxoplasmosisareverydifferent,andmostoftheinfectedpersonsdonotreally feelill.Atypicalsignofthediseaseisaflu-likesensationwithswollenlymphnodesandmuscle pain,whichmaylastformonths.Thesevereformoftoxoplasmosiscausesdamagestothebrain, theeyesandfurtherorgans.Evenwhentheillnesshasceased,areactivechronictypecan develop.Thetransmissionofthediseasetakesprimarilyplacebytheingestionofcatfeces,e.g. duringworkinthegardenorinthecourseofthepetcare.Butinfectiveagentscanalsoappearin rawmeatorcontaminatedwater. Normallytheclinicalsymptomsdisappearaftersomeweeksevenwithouttreatment,butfor pregnantwomenandindividualswithreducedimmunesystematherapywithdrugsshouldbe initiated. Thefollowinglaboratorymethodsareavailable:Complementfixation(CF),immunofluorescence (IFT)orELISA.Thedeterminationofvirus-specificIgMantibodiesinfreshorreactivatedinfections isofspecialimportance,theIgGtestisusedforthedetectionofimmunity.Ifthereisasevere connatalillness,anidentificationoftheparasiteoutofperipheralblood,amnioticliquidortissue samplescanbetried.
3.PrincipleoftheTest TheGenWayToxoplasmaIgGantibodytestkitisbasedontheprincipleoftheenzyme immunoassay(EIA).Toxoplasmaantigenisboundonthesurfaceofthemicrotiterstrips.Diluted sampleserumorready-to-usecalibratorsarePipettedintothewellsofthemicrotiterplate.A bindingbetweentheIgGantibodiesoftheserumandtheimmobilizedToxoplasmaantigentakes place.Afteraonehourincubationatroomtemperature,theplateisrinsedwithdilutedwash solution,inordertoremoveunboundmaterial.Thenready-to-useanti-human-IgGperoxidase conjugateisaddedandincubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB) solutionispipettedandincubatedfor20minutes,inducingthedevelopmentofabluedyeinthe wells.Thecolordevelopmentisterminatedbytheadditionofastopsolution,whichchangesthe colorfrombluetoyellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelength of450nm.TheconcentrationoftheIgGantibodiesisdirectlyproportionaltotheintensityofthe color.