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GenWay/Borrelia burgdorferi IgM recombinant antigens + liquor/GWB-C3EBD6/1x96 Assays
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4000-520-616
Description:
PleaseclickhereforMSDSPDFDatasheetProductName:BorreliaburgdorferiIgMELISA(recombinant)
#ofSamples:1x96Assays
IntendedUse:TherecombinantBorreliaburgdorferiIgM-ELISAisintendedforthequalitativedeterminationofIgMclassantibodiesagainstBorreliaburgdorferiinhumanserumorplasma(citrate)offeringincreaseddiagnosticspecificityandsensitivitybyemployingimmunodominantantigens.IncombinationwiththerecombinantIgGELISAeachclinicalphaseofLymedisease-fromErythmachronicummigransoverBannwarth-SyndromuptoLymearthritis-maybedetectedeveninborderlandcasesprovidingunerringresultsanddiagnosticaidfortheclinicalstaff.
Introduction:SpiRochetesaremotilebacteriawithaperiplasmaticaxialfilament.AllpathogenicspeciesbelongtothefamilyTreponemataceae,whichincludesthethreegenera:Treponema,Borrelia,andLeptospira.TheTreponemaeareextremelylong,flexIBLe,filamentouscellsthatareusuallyheldinacharacteristicspiral,orcoiled-springshape.BorreliaearethelargestTreponemataceaewithverycoarseandirregularspirals.Borreliaburgdorferi,thecausativeagentofLymedisease,istransmittedmainlybyticksbutprobablyalsobyotherblood-suckinginsects.HABItatsarethewooded,humidandtemperateregionsofNorthAmerica,Europe,NorthAfrica,AustraliaandJapan;foresters,farmers,andanybodyenteringinfestedforestsmaybeaffected.
Thedegreeofcontaminationofticksamountsto3-60%dependentonseasonalandregionaldifferences;upto30%ofthepopulationmaybeinfected(about1500casesannuallyinUSA,severalhundredinEurope).
Becauseofitsmultisystemiccharacter,unequivocaldiagnosisofLymediseaseisdifficult.Onlyearlydetectionpermitsefficientcontrolbyantibiotics,sinceinthechronicphaseborreliaearenearlyinaccessible.Thepresenceofbacteriaresp.infectionmaybeidentifiedby:
Microscopy:GiemsaorWright-stainedbloodsmears
PCR
SEROlogy:DetectionofantibodiesbyIF,ELISA,immunoblotting
Principlesoftheassay:ThemajorconstituentofB.burgdorferiflagellaisflagellin(41kDa,p41).WhereasthelipoproteinOspC(22kDa,p22)withintheoutermembraneofthespirocheteinducesanearlyantibodyformationintheECM-phaseofLymedisease.Recombinantp41iisincludedinthepresentedantigenstoavoidcrossreactivitywithantibodiesofsyphiliticsera.
TheBorreliaburgdorferiIgM-ELISAcontainstherecombinantepitopeOspCofthephylumPKo(B.afzelii)and20047(B.garinii)andp41iofthephylumPBi(B.garinii).
Microtiterstripwellsarepre-coatedwithrecombinantBorreliaburgdorferiantigens.DilutedpatientspecimensandreadytousecontrolsareaddedtothesewellsandantibodiesrecognizingtheimmobilizedB.burgdorferiantigenbindduringthefirstincubation.AfterwashingthewellstoremoveallunboundsampleandcontrolmaterialhorserADIshperoxidaselabelledanti-humanIgMconjugateisadded.Duringasecondincubationthisconjugatebindstothecapturedantibodies,andtheexcessunboundconjugateisremovedbyafurtherwashstep.TheimmunecomplexformedbytheboundconjugateisvisualizedwithTMBSubstrateSolutionwhichgivesabluereactionproduct,theintensityofwhichisproportionaltotheamountofB.burgdorferi-specificIgMantibodyinthepatientspecimen.Sulphuricacidisaddedtoeachwelltostopthereaction.Thisproducesayellowendpointcolour.Absorbanceat450nmisreadusinganELISAmicrowellplatereader.
StorageandStability:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.
LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.Diagnosisofaninfectiousdiseaseshouldnotbeestablishedonthebasisofasingletestresult.Aprecisediagnosisshouldtakeintoconsiderationclinicalhistory,symptomatologyaswellasserologicaldata.Inimmunsuppremizedpatientsandnewbornsserologicaldataonlyhaverestrictedvalue.Aneg.result(IgMorIgG)cannotexcludeaninfectionwithB.burgdorferi.Especiallyintheearlyphaseofinfectionthereisthepossibilitythatnoneornodetectionquantitiesoftheantibodiesdoexist.Inthecaseofinfectionoragreyzoneresultwerecommendafurthersampleafter2-3weeks.ApositiveresultofIgMdoesnotalwaysmeanthatanacuteinfectionexistsbecausetheantibodiesofapreviousinfectioncanpersist.Theuseofrecombinantantigensavoidextensivelycrossreactionswiththefollowingantibodies:Treponemapallidum,Leptospira,Borreliarecurrentis.Antibodiesofalues-infectiontop41iwereoccasionallydeterminated.Thereforealues-infectionshouldbeexcluded.
ReferencesJohnsonR.C.,SchmidG.P.,HydeF.W.,SteigerwaldtA.G.,BrennerD.J.(1984)Borreliaburgdorferisp.nov.:etiologicagentofLymedisease.IntJSystBacteriol34,496-497AsbrinkE.,HovmarkA.(1988)EarlyandlatecutaneousmanifestationsofIxodes-bornborreliosis(erythemamigransborreliosis,LymeBorreliose).AnnNYAcadSci539,4-15KarlsonM.,Hovind-HouganK.,SvenungsenB.,StiernstedG.(1990)CultivationandcharacterizationofSpirochetesfromcerebrospinalfluidofpatientswithLymeborreliosis.JClinMicrobiol28,473-479Preac-MursicdV.,WilskeB.,SchierzG.(1986)EurpeanBorreliaBurgdorferiisolatedfromhumanandticks:Cultureconditionsandantibioticsusceptibility.ZblBaktHygA263,112-118WilskeB.,Preac-MursicV.,FuchsR.,SchierzG.(1990)DiagnostikderLymeBorreliose.DiagnoseundLabor,Laboratoriumsblätter40,24-36
AdditionalInformation:
| Name | BorreliaburgdorferiIgMrecombinantantigens+liquor |
|---|---|
| RelatedProductNames | BorreliaburgdorferiIgM–ELISA(recombinant) |
| MolecularWeight | 0.5 |
| Storage | Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C. |
| IntendedUse | ResearchUseOnly |
