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EnzymeimmunoassaybasedonmicrotiterplateforthedetectionandquantitativedeterminationofhumanIgAantibodiesagainstMumpsVirusinserumandplasma.
IntendedUse:
TheMumpsIgAAntibodyELISATestKithasbeendesignedforthethedetectionandthequantitativedeterminationofspecificIgAantibodiesagainstMumpsvirusinserumandplasma.Thisassayisintendedforresearchuseonly.
GeneralInformation:
Mumps(Parotitis)isacommoncontagiousdiseasewithrelativelymoderatesymptomsduringchildhood,butincreasingcomplications,whenadultsareinfected.ThecausativeagentofmumpsisavirusoftheParamyxoviridaefamily.Thevirusnormallyinfectschildrenattheageof4to10.Theinfectionismainlytransmittedbytheairborneroute,butisalsospreadbyvariousobjectscontaminatedbythepatientsaliva.Thediseaseshowsaseasonalprevalencewiththegreatestincidencein
winterandspring.Bothamumpsinfectionorvaccinationleadtoapersistentimmunity.
Thetypicalsymptomassociatedwithamumpsinfectionisa"parotitis“(swellingoftheparotidglands).AdditionallypathologicalinvolvementoftheCNSandvariousglandularorgans(pancreas,thymus,thyroids)isatypicalfeatureofmumps.Amumpsinducedmeningitisisoneofthemostfrequentmanifestationsofthediseasewhichcanalsoappearwithoutparotitis.Thepublisheddataaboutthepresenceofmeningitisvarybetween1.4%and66%oftheclinicallyillpatients.Inmostcasestherefollowsreconvalescencewithoutcomplications.
TherecognitionofMumpsinthelaboratoryismostlydonebydetectionoftheinfectiousagentitselforbythedeterminationofvirus-specificantibodies.TheSEROdiagnosisplaysamajorpart.Besidestheclassicalmethodslikecomplementfixation,hemagglutinationandneutralisationtestshavebeenintroducedaseriesofmodernassayslikeimmunofluorescence,rADIoimmunoassayandenzymeimmunoassay.TheimmunityofanindividualismonitoredbyanIgGELISA.IncreasingIgGtitersarehelpfulinthedeterminationofthecausativeagentbecausecrossreactionswithParainfluenza-Virustype2mayleadtoawronginterpretationofthepatientreports.Intheearlystageofthedisease,significantIgMtitersmaybemonitored.
PrincipleoftheTest:
TheMumpsIgAantibodytestkitisbasedontheprincipleoftheenzymeimmunoassay(EIA).Mumpsantigenisboundonthesurfaceofthemicrotiterstrips.Dilutedpatientserumorready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.AbindingbetweentheIgAantibodiesoftheserumandtheimmobilizedMumpsantigentakesplace.Afteraonehourincubationatroomtemperature,theplateisrinsedwithdilutedwashsolution,inordertoremoveunboundmaterial.Thenready-to-useanti-human-IgAperoxidaseconjugateisaddedandincubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedandincubatedfor20minutes,inducingthedevelopmentofabluedyeinthewells.Thecolordevelopmentisterminatedbytheadditionofastopsolution,whichchangesthecolorfrombluetoyellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelengthof450nm.TheconcentrationoftheIgAantibodiesisdirectlyproportionaltotheintensityofthecolor.