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GenWay/Leishmania infantum IgG (Leishmaniosis)/GWB-A279DD/1x96 Assays

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ProductName:LeishmaniainfantumIgG-ELISA

#ofSamples:1x96Assays

IntendedUse:TheLeishmaniaELISAisintendedforthequalitativedeterminationofantibodiesagainstLeishmaniainfantuminhumanserumorplasma(citrate).

Introduction:Leishmaniaareprotozoabelongingtothefamilytrypanosomatidae.Theparasitesexistintwoforms:thepromastigotesinthemidgutofthevectorinsect,andtheamastigoteswithinthephagolysosomesofmacrophagesintheirmammalianhosts.Inthemacrophagestheyliveasround,non-motileamastigotes(3-7mmindiameter).Themacrophagesareingestedbythesandflyduringblood-mealandtheamastigotesarereleasedintotheirstomach.Almostimmediatelytheamastigotestransformintothemotile,elongated(10-20μm),flagellatepromastigoteform,whichmigratetothealimentarytractoftheflyandaftermultiplicationmoveforwardtothesalivaryglandsoftheinsect.Leishmaniasesareagloballywidespreadgroupofparasiticdiseases;the“type”isdeterminedbytheprimarylocationofthemacrophagesthatareinfected.InhumansfourdifferentformsofLeishmaniasiswithabroadrangeofclinicalmanifestationsarepresent;allcanhavedevastingconsequences.Leishmaniasiscurrentlyaffectssome12millionpeoplein88countries,allbut16ofwhichareinthedevelopingworld.Itisestimatedthat350millionpeopleareexposedtotheriskofinfectionbythedifferentspeciesofLeishmaniaparasite;theannualincidenceofnewcasesisabout2million(1-1.5millioncasesofCL,500,000casesofVL).Visceralleishmaniasis(VL)isthemostsevereformofthedisease,which,ifuntreated,hasamortalityrateofalmost100%.Likemanyothertropicaldiseases,theleishmaniasesarerelatedtoeconomicdevelopmentandman-madeenvironmentalchanges,whichincreaseexposuretothesandflyvector.Thegeographicaldistributionislimitedbythedistributionofthesandfly.AIDSandotherimmunosuppressiveconditionsincreasetheriskofLeishmaniainfectedpeopledevelopingvisceralillness(VL).Leishmania/HIVco-infectionsareconsideredtobeareal“emergingdisease”,especiallyinsouth-westernEurope,where25-70%ofadultVLcasesarerelatedtoHIVinfection,and1.5-9.5%ofAIDScasessufferfromnewlyacquiredorreactivatedVL.Intravenousdrugusershavebeenidentifiedasthemainpopulationatrisk.
Infectionmaybeidentifiedby
Microscopy
SEROlogy:IFA,ELISA

Principlesoftheassay:ThequalitativeimmunoenzymaticdeterminationofantibodiesagainstLeishmaniaisbasedontheELISA(Enzyme-linkedImmunosorbentAssay)technique.MicrotiterstripwellsareprecoatedwithLeishmaniaantigenstobindcorrespondingantibodiesofthespecimen.AfterwashingthewellstoremoveallunboundsamplematerialhorserADIshperoxidase(HRP)labelledProteinAconjugateisadded.ThisconjugatebindstothecapturedLeishmaniaspecificantibodies.TheimmunecomplexformedbytheboundconjugateisvisualizedbyaddingTetramethylbenzidine(TMB)SubstrateSolutionwhichgivesabluereactionproduct.TheintensityofthisproductisproportionaltotheamountofLeishmaniaspecificantibodiesinthespecimen.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolour.Absorbancealternative450nmisreadusinganELISAmicrowellplatereader.

StorageandStABIlity:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.

LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.Diagnosisofaninfectiousdiseaseshouldnotbeestablishedonthebasisofasingletestresult.Aprecisediagnosisshouldtakeintoconsiderationclinicalhistory,symptomatologyaswellasserologicaldata.
Inimmunocompromisedpatientsandnewbornsserologicaldataonlyhaverestrictedvalue.
AcrossreactionwithantibodiesagainstTrypanosomacruzi(Chagas)cannotbeexcluded.

ReferencesWHO.1990.ControloftheLeishmaniases.ReportofaWHOExpertCommittee.Geneva:HealthOrganization,TechnicalReportSeries,No.793.Marsden,P.D.1984.Selectiveprimaryhealthcare:strategiesforcontrolofdiseaseinthedevelopingworld.XIV.Leishmaniasis,Rev.Inf.Dis.6:736-744Ashford,D.a.,Baduro,R.Eulalio,C.,Freire,M.,Miranda,C.,Zalia,M.G.andDavid,J.R.1993.Studiesonthecontrolofvisceralleishmaniasis:validationofthefalconassayscreeningtest-enzyme-linkedimmunosorbentassay(FAST-ELISAa)forfielddiagnosisofcaninevisceralleishmaniasis.Am.J.Med.Hyg.48(1):1-8.Neogy,A.B.,Vouldoukis,A.S.,Otamires,Y.,Tselentis,J.C.,Lascombe,T.,Segalen,D.Rzepka,D.andMonour,L.1993.SerodiagnosisandscreeningofcaninevisceralleishmaniasisinanendemicareaofCorsica:Applicabilityofadirectagglutinationtestandimmunoblotanalysis.AmJTropMedHyg.47:772-777.Evans,T.G.,Vasconcelos,I.A.B.,Lima,J.N.,Teixeira,J.M.,McAullife,I.T.,Lopes,U.G.,Pearson,R.D.,Vasconcelos,A.W.1990.Caninevisceralleishmaniasisinnortheastbrazil:assessmentofserodiagnosismethods.Am.J.Trop.Med.Hyg.42:1118-123.WHO.ReportoftheconsultativeMeetingonHIV/Leishmaniaco-infections.Rome,1994.Allain,D.S.andKagan,I.G.1975.Adirectagglutinationtestforleishmaniasis.Am.J.Trop.Med.Hyg

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NameLeishmaniainfantumIgG(Leishmaniosis)
RelatedProductNamesLeishmaniainfantumIgG-ELISA
MolecularWeight0.5
StorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.
IntendedUseResearchUseOnly