Description:PleaseclickhereforMSDSPDFDatasheetProductName:BordetellapertussisIgM–ELISA#ofSamples:1x96AssaysIntendedUse:TheBordetellapertussisIgM-ELISAisintendedforthequalitativedeterminationofIgMclassantibodiesagainstBordetellapertussisandBordetellapertussistoxininhumanserumorplasma(citrate).Introduction:Bordetellaspeciesarenon-spore-formingencapsulatedbipolar,coccoid(pale-staining)Gram-negativebacilli(about0.3-0.5μmthickand1μmlong).ThegenusconsistsofthehumanparasitesB.pertussisandB.parapertussis,andB.bronchisepticaandB.aviumwhichcauseenzooticinfectionsinvariouswildanddomesticanimalspecies.B.pertussisistheclassicalexciterofpertussisandexistsonlyinillpeople;B.parapertussiscauses5-20%ofamilderandoftenclinicalunapparentformofpertussis.B.bronchisepticahasseldomly(egclosecontactwithanimals)humanpathogenicsignificanceasopportuNISTicsecondaryexciterinmixedinfections(bronchitis,pneumonia,woundinfection).Pertussisorwhoopingcoughisabacterialinfectionoftherespiratorysystem.Itisahighlycontagiouschildhooddiseasewhichappearsseldomlyunderadults.Itistransmittedbyrespiratorycontact.B.pertussishastheABIlitytosticktotheciliaoftheepidermalcellsoftherespiratorysystem.Fragmentsofthebacterialcellwall(ExoToxin=trachealCytotoxine,TCT)inhibitthemovementoftheciliainthetrachealmucosa.Afteranincubationtimeofonetothreeweeksthediseaserunsthroughthreestages(s.tablebelow).Thelethalityis0.6%andconcernsbabiesinthefirstsixmonthswithmorethan70%.Fornewbornandprematureinfantsitishigher(1-2%).InAfricabesidemeaslevirusB.pertussisisthemainreasonforhighinfantmortility.Thedistributionofthediseaseisworldwide.Clinicalpertussisisfollowedbynaturalacquiredimmunitywhichislong-lastingbutnotpermanent.Inmostcountriesanactivevaccinationisrecommendedwhichleadsto90%protectionforthreetotwelveyears.Usuallytheimmunizationpreparationiscombinedwithdiphtheriaandtetanustoxoids.Thepresenceofbacteriumresp.infectionmaybeidentifiedbyMicroscopy:identificationoncultures,IFSEROlogy:DetectionofantibodyproductionbyELISAPrecisediagnosisisneededforeffectivetreatmentofpatients,forisolationofunvaccinatedinfantsatrisk,andfordifferentiationofpertussisfromatypicaldiseasesandchronicinfections.Principlesoftheassay:ThequalitativeimmunoenzymaticdeterminationofIgM-classantibodiesagainstBordetellapertussisisbasedontheELISA(Enzyme-linkedImmunosorbentAssay)technique.MicrotiterstripwellsareprecoatedwithBordetellapertussis/toxinantigenstobindcorrespondingantibodiesofthespecimen.AfterwashingthewellstoremoveallunboundsamplematerialhorserADIshperoxidase(HRP)labelledanti-humanIgMconjugateisadded.ThisconjugatebindstothecapturedBordetellapertussis/toxinspecificantibodies.Theimmunecomplexformedbytheboundconjugateisvisualizedbyaddingtetramethylbenzidine(TMB)substratewhichgivesabluereactionproduct.TheintensityofthisproductisproportionaltotheamountofBordetellapertussis/toxinspecificIgMantibodiesinthespecimen.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolour.Absorbanceat450nmisreadusinganELISAmicrowellplatereader.StorageandStability:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2…8°C.LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.Diagnosisofaninfectiousdiseaseshouldnotbeestablishedonthebasisofasingletestresult.Aprecisediagnosisshouldtakeintoconsiderationclinicalhistory,symptomatologyaswellasserologicaldata.InimmunsuppremizedpatientsandnewbornsserologicaldataonlyhaverestrictedvalueReferencesWong,K.H.,andS.K.Skelton.1989.PreparationoffilamentoushemagglutininfromBordetellapertussisandassayforserumantibodiestofilamentoushemagglutininandpertussistoxinforclinicalandpublichealthlab.J.Clin.Microbiol.27:2805-2810Zackrission,G.,I.,Krantz,T.Legergard,P.Larsson,R.Sekura,N.Sigurs,J.Taranger,andB.Trollfors.1988.HumoralantibodyresponsetopertussistoxininpatientswithclinicalpertussisZachrisson,G.,I.Krantz,T.Lagergard,P.Larsson,R.Sekura,NSigurs,S.Taranger,andB.Trollfors.1988.Humoralantibodyresponsetopertussistoxininpatientwithclinicalpertussismeasuredbyanenzyme-linkedimmunosorbentassay.Eur.J.Clin.Microbiol.7:149-154Granstrom,M.,G.Granstrom,A.Lindtors1982.Serologicdiagnosisofwhoopingcoughbyanenzyme-linkedimmunsorbentassayusingfimbrialhemagglutininasantigen.J.Infect.Dis.146:741-745Wong,K.H.,andS.K.Skelton.1989.PreparationoffilamentoushemagglutininfromBordetellapertussisandassayforserumantibodiestofilamentoushemagglutininandpertussistoxinforclinicalandpublichealthlab.J.Clin.Microbiol.27:2805-2810Zackrission,G.,I.,Krantz,T.Legergard,P.Larsson,R.Sekura,N.Sigurs,J.Taranger,andB.Trollfors.1988.HumoralantibodyresponsetopertussistoxininpatientswithclinicalpertussisZachrisson,G.,I.Krantz,T.Lagergard,P.Larsson,R.Sekura,NSigurs,S.Taranger,andB.Trollfors.1988.Humoralantibodyresponsetopertussistoxininpatientwithclinicalpertussismeasuredbyanenzyme-linkedimmunosorbentassay.Eur.J.Clin.Microbiol.7:149-154Granstrom,M.,G.Granstrom,A.Lindtors1982.Serologicdiagnosisofwhoopingcoughbyanenzyme-linkedimmunsorbentassayusingfimbrialhemagglutininasantigen.J.Infect.Dis.146:741-745AdditionalInformation:NameBordetellapertussisIgMRelatedProductNamesBordetellapertussisIgM–ELISAMolecularWeight0.5StorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2…8°C.IntendedUseResearchUseOnly