Description:ProductNameMycoplasmapneumoniaeIgM-ELISA#ofSamples1x96Assays.IntendedUseTheMycoplasmapneumoniaeIgM-ELISAisintendedforthequalitativedeterminationofIgMclassantibodiesagainstM.pneumoniaeinhumanserumorplasma(citrate).IntroductionThemycoplasmsbelongtotheclassMollicutescomprisingthreedistinctfamiliesandfourgenera,oneofwhichisMycoplasmawithover60species.Mycoplasmaearethesmallestfreelivingorganismsknown(300to500nmindiameter)andunlikeregularbacteriatheylackacellwall.Mycoplasmsareextracellularparasites,especiallyonmucousmembranes,whichcancauseinfectionsinhu-mans,animals,plants,andcellcultures.Mycoplasmapneumoniaeisprimarilyarespiratorypathogen(obligat)inhumansinvolvingthenasopharynx,throat,trachea,bronchi,bronchioles,andalveoli.OtherMycoplasmae,M.buccale,M.faucium,M.oraleandM.salivariumarecommensalsintheoralcavity.MycoplasmahominisandUreaplasmaurealyticuminhABItprimarilythegenitaltractandmayactasopportuNISTicinvaders.M.pneumoniaeisbyfarthemostimportantpathogenofthisgroup.InfectionwithM.pneumoniaeoccursworldwide,itsepidemiologyhasbeenstudiedprimarilyintheUSA,Europe,andJapan.Infectionsareendemicinlargerurbanareas,andepidemicincreasesareobservedatvaryingintervalls.Mycoplasmapneumoniaehasbeenestimatedtocause15-20%ofallpneumoniae;therateishighestinchildrenandyoungadults.74%ofinfectionswithM.pneumoniaeareasymptomatic,reinfectionmayoccur.NaturallyacquiredimmunitytoinfectionwithM.pneumoniaeappearstobeoflimitedduration(2-3years).InfectionmaybeidentifiedbyMicroscopyHemadsorption,Tetrazoliumreduction(specificforM.pneumoniae),ComplementfixationDetectionofantibodyproductionbyELISASEROlogy:complementfixation(CF),neutralization(N)andhemagglutination-inhibition(HAI);DetectionofantibodiesandthehexonantigenbyELISA.PrinciplesoftheassayThequalitativeimmunoenzymaticdeterminationofIgM-classantibodiesagainstMycoplasmapneumoniaeisbasedontheELISA(Enzyme-linkedImmunosorbentAssay)technique.MicrotiterstripwellsareprecoatedwithMycoplasmapneumoniaeantigenstobindcorrespondingantibodiesofthespecimen.AfterwashingthewellstoremoveallunboundsamplematerialhorserADIshperoxidase(HRP)labelledanti-humanIgMconjugateisadded.ThisconjugatebindstothecapturedMycoplasmapneumoniae-specificantibodies.TheimmunecomplexformedbytheboundconjugateisvisualizedbyaddingTetramethylbenzidine(TMB)substratewhichgivesabluereactionproduct.TheintensityofthisproductisproportionaltotheamountofMycoplasmapneumoniae-specificIgMantibodiesinthespecimen.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolour.Absorbanceat450nmisreadusinganELISAmicrowellplatereader.StorageandStabilityThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2-8°C.LimitationsoftheTestBacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.Diagnosisofaninfectiousdiseaseshouldnotbeestablishedonthebasisofasingletestresult.Aprecisediagnosisshouldtakeintoconsiderationclinicalhistory,symptomatologyaswellasserologicaldata.Inimmunocompromisedpatientsandnewbornsserologicaldataonlyhaverestrictedvalue.ReferencesClydeWAJr:ClinicaloverviweoftypicalM.pneumoniaeinfections.Clin.Infect.Dis1993Aug;17Suppl1:S32-6MedlineE.Jacobs.1993.SerologicalsDiagnosisofM.peumoniaeInfections:CurrentProdedures.ClinicalInfectiousDiseases.17(suppl1)79-82Foy,H.M.1993.InfectionscausedbyM.pneumoniaeandpossIBLepopulationsofpatients.ClinicalInfectiousDiseases.17(suppl1)37-46HandleyJ.G.,andL.D.Gray.1997.TheincidenceofM.pneumoniaeBoardFamilyPractice11(6):425-429Vikerfors,T.,Brodin,G.,Grandien,M.,Hirshberg,L.,Krook,A.,andC.A.PetterssonspecificIgMantibodiesforthediagnosisofM.pneumoniaeinfectionsScand.J.Infect.Dis.20(6):601-610Wiegand,R.:M.pneumoniae.In:Brandis,H.,W.Kohler,H.J.Eggers,G.Pulverer(ed.):Med.MikroBIOLOGie1994.GustavFischerVerlagStuttgart,Jena,NewYork(1994)767-769.AdditionalInformation:NameMycoplasmapneumoniaeIgM-ELISARelatedProductNamesMycoplasmapneumoniaeIgM-ELISAStorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.IntendedUseResearchUseOnly