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1.IntendedUse TheGenWayHerpes1IgAAntibodyELISATestKithasbeendesignedforthethedetectionand thequantitativedeterminationofspecificIgAantibodiesagainstHerpes1inserumandplasma. FurtherapplicationsinotherbodyfluidsarepossIBLeandcanberequestedfromtheTechnical ServiceofGENWAY. Thisassayisintendedforresearchuseonly.
2.GeneralInformation TheHerpessimplexvirustype1isanubiquitouspathogenofhumansthatusuallycauseseitherasymptomaticinfectionormildskinandmucosaldiseases.AntibodiestoHSV1occurinabout90% ofadults.NormallyHSV1istransmittedbyoralsecretionsoropenwoundspriortotheageoffive. Recentlyinadultsprimaryinfectionswereobserved,too.Aftertheprimaryinfectionsomeviruses establishalatentstateintheirhostcells(mostlyganglialcells).ThevirusDNAisintegratedinto thegenomeofthehostcell,whereitremainsuntiltheinfectedpersondies.Afterstimulationofthe hostcell,recurrentinfectionoccurs,whichiscalledanexacerbation,whenclinicalsymtomsappear. Therecurrencemaybecausedbydifferentkindsoftraumas,asfeverorphysiologicalchangesand diseases.Immunosuppressedpersonsmayshowasevereclinicalcourse.HSV1causesdifferent clinicalsymptomsinabout10%oftheprimaryinfections.HSV1causes85%andHSV215%of oralprimaryinfections.ThemajorclinicalmanifestationsassociatedwithHSV1infectionsare gingivostomatitis,keratitis,conjunctivitis,vesiculareruptionsoftheskin,encephalitis,eczemaand somelethalinfectionsofnewborns.Insome casesHSV1infectionleadstoameningitiswith differentneurologicalsymptoms.PersonsatanincreasedriskforseriousorprolongedHSV infectionsarethosewitheczema,severeburnsoradefectintheircell-mediatedimmunity.Thedrug AcycloviristhetreatmentofchoiceformostseriousHSVinfections.Thecommonmanifestations ofHSVinfectionsaresotypicalthattheinfectionmaybeeasilydiagnosedonclinicalrecognition alone.The"goldstandard"fordiagnosisofHSVinfectionremainsisolationofthevirusintissue culture.FortypingHSV1/HSV2Westernblotsorindirectimmunofluorescencemaybeperformed. DiagnosisoftheprimaryinfectionbyHSV1/HSV2canbeconfirmedbyasignificantriseofthe IgGtiterwithin6to10days.AfinishedinfectioncanbemonitoredbytheIgGELISA.Incaseofa sUSPicionofHSVencephalopathyitisrecommendedtoperformaparalleldeterminationofboth HSV-specificantibodies(IgGandIgM)inserumandliquor.
3.PrincipleoftheTest TheGENWAYHerpes1IgAantibodytestkitisbasedontheprincipleoftheenzyme immunoassay(EIA).Herpes1antigenisboundon thesurfaceofthemicrotiterstrips.Diluted sampleserumorready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.Abinding betweentheIgAantibodiesoftheserumandtheimmobilizedHerpes1antigentakesplace.Aftera onehourincubationatroomtemperature,theplateisrinsedwithdilutedwashsolution,inorderto removeunboundmaterial.Thenready-to-useanti-human-IgAperoxidaseconjugateisaddedand incubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedand incubatedfor20minutes,inducingthedevelopmentofabluedyeinthewells.Thecolor developmentisterminatedbytheadditionofastopsolution,whichchangesthecolorfromblueto yellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelengthof450nm.The concentrationoftheIgAantibodiesisdirectlyproportionaltotheintensityofthecolor.