Description:1.IntendedUse TheGenWayAspergillusfumigatusIgGAntibodyELISATestKithasbeendesignedforthethe detectionandthequantitativedeterminationofspecificIgGantibodiesagainstAspergillus fumigatusinserumandplasma.FurtherapplicationsinotherbodyfluidsarepossIBLeandcanbe requestedfromtheTechnicalServiceofGenWay. Thisassayisintendedforresearchuseonly. 2.GeneralInformation AspergillusspeciesofknownpathogenicitytomanareAspergillusfumigatus,A.flavus,A.niger,A. terreusandA.nidulans.ThemostcommonpathogenofthisgenusisA.fumingatuswhichoccursin hay,grain,rottenplantsandbirdsfaeces.ThemainopportuNISTicinvasivefungialinfectionsarethe candidalmycosisfollowedbyaspergillosis.GenerallyinfectionswithAspergillusspp.areairborne. BecauseoftheubiquityofAspergillusspecies itrendersmoredifficulttodecidebetween contaminationbycommensalsoraseriousinfection.Usuallyinfectioninmanoccursinalready damagedtissuesonly.Aspergillusspp.cancauseachronicalinfectionofparanasalsinus,eyesor lungs. Threetypesoflung-aspergillosiscanbedistinguished: a:Acuteinfection(bronchialpneumonia;pneumonia)Aspergilluspneumoniaismostlyfoundinindividualswithneutropenia(decreaseofneutrophil granulocytes),afteralong-timetherapywith glucocorticoids,inimmunosuppressedindividuals (afterorgantransplantation)andinalcoholics. b:Saprophyticaspergillom(compactreticulumofhyphaeinthelungs)Preformedcavesinthelungduetoaprevious tuberculosisgiveplacetoacolonisationof Aspergillusspecies. c:AllergicbronchopulmonalaspergillosisThispictureisnotduetoaninfectiousdiseasebutahypersensitivereactionofthebronchialsystem (mediatedbyIgE)afterinhalationofaspergillusspores.Subsequentlythebronchialsystem produceshighlyviscoussecretions,thatmayblockthebronchiallumen.Theindividualdevelops difficultiesofbreathingandafibrosis. NexttoELISAtheindirectAspergillushemagglutinationtest(AspergillusHAT)canbeperformed todetectspecificIgGandIgMantibodies.TheHATisnotsuitableasascreeningtest,however, becauseofitslowsensitivity.Insomehigh-riskindividualsitshowsonlylowantibodytiters.Fora betterdiagnosisofinvasiveaspergillosisthebrainorlungoftheseindividualsshouldbeexamined byabiopsy. 3.PrincipleoftheTest TheGenWayAspergillusfumigatusIgGantibodytestkitisbasedontheprincipleoftheenzyme immunoassay(EIA).Aspergillusantigenisboundonthesurfaceofthemicrotiterstrips.Diluted sampleserumorready-to-usestandardsarePipettedintothewellsofthemicrotiterplate.Abinding betweentheIgGantibodiesoftheserumandtheimmobilizedAspergillusantigentakesplace.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:NameAspergillusfumigatusIgGELISARelatedProductNamesAspergillusfumigatusIgGELISAApplicationsEnzymeimmunoassaybasedonmicrotiterplateforthedetectionandquantitativedeterminationofhumanIgGantibodiesagainstAspergillusfumigatusinserumandplasmaIntendedUseResearchUseOnly