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ProductName:HelicobacterpyloriIgGplus–ELISA

#ofSamples:1x96Assays

IntendedUse:TheHelicobacterpyloriIgGplus-ELISAisintendedforthequalitativeandquantitativedeterminationofIgGclassantibodiesagainstHelicobacterpyloriinhumanserumorplasma(citrate).

Introduction:HelicobacterpyloriisaspiralGram-negativebacterium(2-6.5μminsize,flagellated)whichcolonizesthehumangastricmucosa.Theorganismisfoundinthemucouslayerandadherestothesurfacemucousepitheliumofthestomachbutgenerallydoesnotpenetratethegastricmucosadirectly.
However,thereisasecondaryinflammatoryresponseinthemucosaleADIngtochronicactivegastritis.Helicobacterpyloriistheprimarycausativeagentinmostcasesofpepticulcerdisease.InfectionrateinEuropeisabout30%-40%,worldwideabout50%.ThereisaninverserelationshipbetweenthepresenceofHelicobacterpyloriinfectionandsocioeconomicstatus.Indevelopingcountries,peopleacquiretheinfectionatanearlyagesuchthatbyyoungadulthoodasmanyas90%ofthepopulationmighthaveHelicobacterpylorigastritis.IndevelopedwesterncountriestheprevalenceofHelicobacterpylorigastritisismuchlower.Undertheseconditions,therateofacquisitionismuchslower(roughly1%perannum)andtheolderoneis,themorelikelyoneistobeinfectedwiththeorganism.
Infectionmaybeidentifiedby
Histology:Giemsa,WarthinStarryorGentastain/cultureofantralbiopsyspecimens
Enzymology:Detectionofbacterialurease(ureabreathtest)
SEROlogy:Detectionofantibodies

Principlesoftheassay:ThequantitativeimmunoenzymaticdeterminationofIgG-classantibodiesagainstHelicobacterpyloriisbasedontheELISA(Enzyme-linkedImmunosorbentAssay)technique.
MicrotiterstripwellsareprecoatedwithHelicobacterpyloriantigenstobindcorrespondingantibodiesofthespecimen.Afterwashingthewellstoremoveallunboundsamplematerialhorseradishperoxidase(HRP)labelledanti-humanIgGconjugateisadded.ThisconjugatebindstothecapturedHelicobacterspecificantibodies.TheimmunecomplexformedbytheboundconjugateisvisualizedbyaddingTetramethylbenzidine(TMB)substratewhichgivesabluereactionproduct.
TheintensityofthisproductisproportionaltotheamountofHelicobacterspecificIgGantibodiesinthespecimen.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolour.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.PyloriandgastricMALTlymphomaLancet339:745-746

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Name	HelicobacterpyloriIgGplus
RelatedProductNames	HelicobacterpyloriIgGplus–ELISA
MolecularWeight	0.5
Storage	Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.
IntendedUse	ResearchUseOnly