
Description:
1.INTRODUCTION
Anti-neutrophilic-cytoplasmantibodies(ANCA)representsagroupofautoantibodiesdirectedtowardsthecytoplasmaticcomponentsoftheneutrophilicgranulocytesandmonocytes.TheclassicalmethodsforthedeterminationofANCAaretheimmunofluorescentmethods.Withtheseindirectimmunofluorescencetechniquestwomainpatternsarerecognized,acytoplasmatic(c-ANCA)andaperinuclear(p-ANCA)type.Themainantigenforthec-ANCAistheproteinase3(PR3),whichisaserineproteinasepresentinprimarygranules.Antibodiesofp-ANCApositiveseraaremainlydirectedtomyeloperoxidase(MPO).Antibodiestootherantigense.g.lactoferrin,elastase,cathepsin-Gandalsolysozymeoftenresultinasimilarp-ANCApattern.Besidedifferentuntypicalvariantsofp-ANCAIFpatternsgranulocytespecificantinuclearantibodies(GS-ANA)isindistinguishablefromp-ANCA.ThismakesaclearinterpretationandclassificationoftheIFpatternsdifficult.ThereforeeverypositiveIF-ANCAfindingsesp.p-ANCAshouldbedifferenciatedbyELISAtechniquesusingpurifiedantigens.AsurveyofdocumentedclinicalindicationsofspecificANCAisgiveninthetablebelow.PR3-ANCAandMPO-ANCAarereliableSEROlogicMarkersinthediagnosticsofvasculitides.PR3-ANCAistheclassicalautoantigeninWegener´sgranulomatosiswithaclinicalspecificityofmorethan95%.c-ANCAisdocumentedtobepresentindifferentdiseases.Anti-MPOantibodiesarehighlyspecificforidiopathicandvasculitisassociatedcrescenticglomerulonephritisandalsoforclassicpolyarteritisnodosa,Churg-Strausssyndromeandthepolyangitisoverlapsyndromewithoutrenalinvolvement.Withrespecttosensitivity,eitherMPOorPR-3antibodieswerefoundin77to100%ofpatientswithidiopathicandvasculitisassociatedcrescenticglomerulonephritis.InWG,anti-MPOantibodiesweredetectedonlyoccasionallyandgenerallyinpatientsnegativeforPR-3antibodies.TheMPOandPR-3specificELISAmethodscanprovideanimportantconfirmatoryresultfortwoofthemoreimportantoftheidentifiedantigens.ELISAisalsousefulforinterpreting"difficult"samplesbyIFAsuchasthosewhichexhibitseveralantibodiessimultaneouslyorthosewithhighbackgroundfluorescence.
2.INTENDEDUSE
AntiPR3isanindirectsolidphaseenzymeimmunoassay(ELISA)forthequantitativemeasurementofIgGclassautoantibodiesagainstproteinase3(PR3)inhumanserumorplasma.Theassayisintendedforlaboratoryresearchuseonly.AntiPR3(c-ANCA)kitisintendedforlaboratoryuseonly.
3.PRINCIPLEOFTHEASSAY
Theanti-PR3IgGtestisbasedonthebindingofpresentantibodiesonthehumanneutrophilproteinase3coatedintothemicroplates.Anypresentantibodiesincalibrators,controlsorpre-dilutedpatientsamplesbindtotheinnersurfaceofthewells.Aftera60minutesincubationthemicroplateiswashedwithwashbufferforremovingnon-reactiveSerumorPlasmacomponents.Ananti-human-IgGhorserADIshperoxidaseconjugatesolutionrecognizeIgGclassantibodiesboundtotheimmobilizedantigens.Aftera30minutesincubationanyexcessenzymeconjugate,whichisnotspecificallyboundiswashedawaywithwashbuffer.AchromogenicsubstratesolutioncontainingTMBisdispensedintothewells.After15minutesofincubationthecolordevelopmentisstoppedbyaddingthestopsolution.Thesolutioncolorchangeintoyellow.TheamountofcolorisdirectlyproportionaltotheconcentrationofIgGantibodiespresentintheoriginalsample.
AdditionalInformation:
Name | AntiPR3(c-ANCA) |
---|---|
RelatedProductNames | Cytoplasmicantineutrophilcytoplasmicantibodies(c-ANCA)ELISA,CytoplasmicantineutrophilcytoplasmicantibodiesELISA,c-ANCAELISA,AntiPR3ELISA,ANCA/VasculitisAssay |
MolecularWeight | 0.5 |
Concentration | LotSpecific |
Applications | EnzymeimmunoassayforthequantitativedeterminationofantiPR3inhumanserumorplasma |
Reactivity | Human |
Storage | Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°Cinthedark. |
IntendedUse | ResearchUseOnly |