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GenWay/Toxoplasma gondii IgM μ-capture– ELISA/GWB-990FCF/1x96 Assays

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ProductName:ToxoplasmagondiiIgMμ-capture–ELISA

#ofSamples:1x96Assays

IntendedUse:TheToxoplasmagondiiIgMμ-captureELISAisintendedforthequalitativedeterminationofIgMclassantibodiesagainstToxoplasmagondiiinhumanserumorplasma(citrate).

Introduction:Toxoplasmagondiiisasmallintracellularparasite,whoselivecyclehasasexualandanasexualphase.Sexualdevelopmentisrestrictedtotheintestinalcellsof(probablyexclusively)cats;theoocystsformedareexcretedandduetotheirresistantcellwallstheymaybeinfectiousunderadvantageouscircumstancesforatleast1year.Animalsandmanareintermediatehostsforthe
asexualproliferationofT.gondii:theingestedparasiteswillproliferateexplosivelywithinthehostcellslysingthemeventually.Theydisseminatethroughoutthebodyviacirculationandlymphaticsystemandthoughmayinfectanycelltype.Inmuscleandbraincellscystsareformedwhicharespheroidalandabout5-100μmindiameter.Cystsarevirtuallyimmortalintheintermediatehost.
Toxoplasmagondiiisthemostcommonparasiteinhumans,butitsabundance(7-80%)ishighlydependentonthegeographicarea,thesocioeconomicstatusandthenutritionalcustoms.Infectiononlyrarelycausestoxoplasmosisandusuallyclinicalsymptomsareabsent,butmayproducesevereproblemsinimmunosuppressedpersonsandfetus.
Becauseonlyaprimaryinfectionduringpregnancymaybedangerousandevenfatalfortheunborn(theprobABIlityofcongenitalinfectionisabout50%),therecentonsetofaninfectionmustbeexcluded.
Inpregnantwomeninover98%ofcases,theabsenceofIgMexcludesthepossibilityofrecentinfection.Innewbornstheverypresenceofanti-toxoplasmaIgMissufficienttoconfirmacongenitaltoxoplasmosis,sincematernalIgM,unlikeIgG,doesnotcrosstheplacentalbarrier.ButasignificantnumberofinfectedinfantsdonotdevelopdetectableIgMlevelsandthusarefalsenegative.Inimmunosuppressedpatientstoxoplasmosiscausesseverecomplicationsmostlybyreactivationofanearlierlatentinfection.Infectionmaybeidentifiedby:
PCR
Indirectimmunofluorescence(IIF)
SEROlogy:DetectionofantibodyproductionbyELISA

Principlesoftheassay:ThequalitativeimmunoenzymaticdeterminationofIgM-classantibodiesagainstToxoplasmagondiiisbasedontheELISA(Enzyme-linkedImmunosorbentAssay)technique.
TheToxoplasmaIgMELISAisanIgM-μ-captureELISA.
Microtiterstripwellsareprecoatedwithanti-humanIgM-classantibodiestobindcorrespondingantibodiesofthespecimen.AfterwashingthewellstoremoveallunboundsamplematerialacomplexofantigenandhorserADIshperoxidaselabelledToxoplasmagondiiantibodyisadded.ThiscomplexbindstothecapturedToxoplasmaspecificIgMantibodies.ThisimmunecomplexisvisualizedwithTetramethylbenzidine(TMB)substratewhichgivesabluereactionproduct.
TheintensityofthisproductisproportionaltotheamountofToxoplasmagondii-specificIgMantibodyinthespecimen.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolour.Absorbanceat450nmisreadusinganELISAmicrowell
platereader.

StorageandStability:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.

LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.Diagnosisofaninfectiousdiseaseshouldnotbeestablishedonthebasisofasingletestresult.Aprecisediagnosisshouldtakeintoconsiderationclinicalhistory,symptomatologyaswellasserologicaldata.
Inimmunocompromizedpatientsandnewbornsserologicaldataonlyhaverestrictedvalue.
Toestimate(primaryorrecurrent)T.gondiiinfectionsbyserologyitisadvisedtotestserumpairs.Thesecondpaircanbedrawn14to21daysafterthefirst.Eachserumpairshouldbetestedatthesamedayandinthesametesttoallowinterpretationofsignificantdifferencesinantibodylevels.ItisadvisedtoperformacombinationofIgM,IgAandIgGtesting.

ReferencesA.Balsarietal.ELISAforToxoplasmaantibodydetection:acomparisionwithotherserodiagnostictests.J.Clin.Pathol.,33:640(1980)M.H.Beaman,R.E.McCabe,S-Y.Wong,J.S.Remington,Toxoplasmagondii,Inc:PrinciplesandPracticeofInfectiousDiseases,G.L.Mandell,J.E.Bennet,R.Dolin,eds.,ChurchillLivingstonePubl.,Fourthedition,p.2455-2475(1995)A.Bondioloetal.ELISAforToxoplasmaantibodies.Inc:ImmunoenzymeAssayTechniques,R.Malvanoed.,M.NijhoffPubl.(1980)M.E.Camargoet.al.ImmunoglobineGandimmunoglobulinMenzyme-linkedimmunosorbentassayanddefinedtoxoplasmosisserologicalpatterns.Inf.Immun.,21:55(1978)Y.Carlieretal.Evaluationoftheenzyme-linkedimmunosorbentassay(ELISA)andotherserologicaltestsforthediagnosisoftoxoplasmosis.Bull.WorldHealthOrganization,58(1):99(1980)

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NameToxoplasmagondiiIgMμ-capture–ELISA
RelatedProductNamesToxoplasmagondiiIgMμ-capture–ELISA
StorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.
IntendedUseResearchUseOnly