Description:PleaseclickhereforMSDSPDFDatasheetProductName:HelicobacterpyloriIgAplusELISA#ofSamples:1x96AssaysIntendedUse:TheHelicobacterpyloriIgAplus-ELISAisintendedforthequantitativedeterminationofIgAclassantibodiesagainstHelicobacterpyloriinhumanserumorplasma(citrate).Introduction:HelicobacterpyloriisaspiralGram-negativebacterium(2-6.5μminsize,flagellated)whichcolonizesthehumangastricmucosa.Theorganismisfoundinthemucuslayerandadherestothesurfacemucusepitheliumofthestomachbutgenerallydoesnotpenetratethegastricmucosadirectly.However,thereisasecondaryinflammatoryresponseinthemucosaleADIngtochronicactivegastritis.Helicobacterpyloriistheprimarycausativeagentinmostcasesofpepticulcerdisease.InfectionrateinEuropeisabout30%-40%,worldwideabout50%.ThereisaninverserelationshipbetweenthepresenceofHelicobacterpyloriinfectionandsocioeconomicstatus.Indevelopingcountries,peopleacquiretheinfectionatanearlyagesuchthatbyyoungadulthoodasmanyas90%ofthepopulationmighthaveHelicobacterpylorigastritis.IndevelopedwesterncountriestheprevalenceofHelicobacterpylorigastritisismuchlower.Undertheseconditions,therateofacquisitionismuchslower(roughly1%perannum)andtheolderoneis,themorelikelyoneistobeinfectedwiththeorganism.InfectionmaybeidentifiedbyHistology:Giemsa,WarthinStarryorGentastain/cultureofantralbiopsyspecimensEnzymology:Detectionofbacterialurease(ureabreathtest)SEROlogy:DetectionofantibodiesPrinciplesoftheassay:ThequantitativeimmunoenzymaticdeterminationofIgA-classantibodiesagainstHelicobacterpyloriisbasedontheELISA(Enzyme-linkedImmunosorbentAssay)technique.MicrotiterstripwellsareprecoatedwithHelicobacterpylorirecombinantantigenstobindcorrespondingantibodiesofthespecimen.Afterwashingthewellstoremoveallunboundsamplematerialhorseradishperoxidase(HRP)labelledanti-humanIgAconjugateisadded.ThisconjugatebindstothecapturedHelicobacter-specificantibodies.TheimmunecomplexformedbytheboundconjugateisvisualizedbyaddingTetramethylbenzidine(TMB)substratewhichgivesabluereactionproduct.TheintensityofthisproductisproportionaltotheamountofHelicobacter-specificIgAantibodiesinthespecimen.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolor.Absorbanceat450nmisreadusinganELISAmicrowellplatereader.StorageandStABIlity:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.Diagnosisofaninfectiousdiseaseshouldnotbeestablishedonthebasisofasingletestresult.Aprecisediagnosisshouldtakeintoconsiderationclinicalhistory,symptomatologyaswellasserologicaldata.Inimmunocompromizedpatientsandnewbornsserologicaldataonlyhaverestrictedvalue.ReferencesMarshall,B.J.,Royce,H.,D.I.,Goodwin,C.S.,Taylor,N.S.,Edmonds,P.,Sly,L.I.,Brenner,D.J.(1982)OrginalIsolationofCampylobacterpyloridisfromhumangastricmucosa.MicrobiosLetter25:83WarrenJ.R.(1983)Unidentifiedcurvedbacilliongastricepitheliuminactivechronicgastritis,Lancet:1273-1275Parsonnet,J.,Friedman,G.D.,Vanderstetten,D.P.,Chang,y.,Vogelman,J.H.,Orentreich,N.,SIBLey,R.K.(1991)H.Pyloriinfectionandtheriskofgastriccarcinoma.NEnglJMed325:1127-1131Stolte,M.Eidt,S.(1993)HealinggastricMALTlymphomasbyeradicatingH.PyloriLancet342:568Stolte,M.(1992)H.PyloriandgastricMALTlymphomaLancet339:745-746AdditionalInformation:NameHelicobacterpyloriIgAplusRelatedProductNamesHelicobacterpyloriIgAplusELISAMolecularWeight0.5StorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.IntendedUseResearchUseOnly