Description:PleaseclickhereforMSDSPDFDatasheetProductName:BrucellaIgG–ELISA#ofSamples:1x96AssaysIntendedUse:TheBrucellaIgG-ELISAisintendedforthequalitativedeterminationofIgGclassantibodiesagainstBrucellainhumanserumorplasma(citrate).Introduction:BrucellaisasmallGram-negativebacterium(0.4-0.8μmindiameterand0.4-3.0μminlength)whichisnon-flagellated,andnon-spore-forming.TheyarenamedafterthemilitarydoctorDavidBruce,whoonMaltain1887isolatedthepathogensfromthespleenofasoldierwhodiedofundulatingfever.Fourspeciesarepathogenictohuman:Brucellaabortus,Brucellamelitensis,BrucellasuisandBrucellacanis.AllfourspeciesareexcitersofBrucellosis,adiseasecharacterizedbyundulatingfever.DependingonexciterthediseaseisalsocalledMorbusBang(B.abortus)orMaltafever(B.melitensis).Thepathogensaretransmittedfromanimals,whicharemainlyaffected.Theinfectioniscausedbycontactwithillanimalsortheirexcrementsaswellasbynon-pasteurizedmilkandmilkproductslikefreshcheesefromsheeporgoat.Mainentrancesareskinwounds,conjunctivesanddigestivetract.Theintactpathogensaretransportedbygranulocytesintolocallymphnodes,fromwheretheyspreadhaematogenous.Allkindoforganscanbeinfected.Symptomsdependupontheinfectedorgan,whereaninflammationtakesplace.Untilnowthepathogenicmechanismisnotcompletelyunderstood.Brucellosisappearsworldwide.Innon-pasteurizedmilkandmilkproductsBrucellaisviablyandinfectiousforweeks.BovinebrucellosiscausedbyBrucellaabortusisstillthemostwidespreadform,althoughreportedincidenceandprevalenceofthediseasevarywidelyfromcountrytocountry(from200per100,000population).Brucellamelitensisisendemicinareaswherekeepingofsheepsandgoatsisfrequent.Itcausesserioushumaninfections.Workingwiththesebacteriainlaboratoriesnecessitateshighestcarefulnessbecauseofhighcontagiosity.Riskgroupsincludeabattoirworkers,meatinspectors,animalhandlers,veterinarians,andlaboratorians.Brucellosisisanationallynotifiablediseaseandreportabletothelocalhealthauthority.ThepresenceofinfectionmaybeidentifiedbyHistology:BrucellaspeciesmaybespeciatedbysusceptibilitytothedyesfuchsinandthioninSEROlogy:DetectionofantibodiesbyELISAPrinciplesoftheassay:ThequalitativeimmunoenzymaticdeterminationofIgG-classantibodiesagainstBrucellaisbasedontheELISA(Enzyme-linkedImmunosorbentAssay)technique.MicrotiterstripwellsareprecoatedwithBrucellaantigenstobindcorrespondingantibodiesofthespecimen.AfterwashingthewellstoremoveallunboundsamplematerialhorserADIshperoxidase(HRP)labelledanti-humanIgGconjugateisadded.ThisconjugatebindstothecapturedBrucella-specificantibodies.TheimmunecomplexformedbytheboundconjugateisvisualizedbyaddingTetramethylbenzidine(TMB)substratewhichgivesabluereactionproduct.TheintensityofthisproductisproportionaltotheamountofBrucella-specificIgGantibodiesinthespecimen.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolour.Absorbanceat450nmisreadusinganELISAmicrowellplatereader.StorageandStABIlity:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.Diagnosisofaninfectiousdiseaseshouldnotbeestablishedonthebasisofasingletestresult.Aprecisediagnosisshouldtakeintoconsiderationclinicalhistory,symptomatologyaswellasserologicaldata.Inimmunocompromisedpatientsandnewbornsserologicaldataonlyhaverestrictedvalue.ReferencesSALATA,R.A.,RAVDIN,J.I.(1985)BRUCELLASPECIES(BRUCELLOSIS).IN:MANDELGL,DOUGLASRG,BENNETJE,EDS.PRINCIPLESANDPRACTICEOFINFECTIOUSDISEASES.NEWYORK:JOHNWILEY,1283-90ARIZA,J.,PELLICER,T.,PALLARÉS,R.,FOZ,A.ANDGUDIOL,F.(1992)SPECIFICANTIBODYPROFILEINHUMANBRUCELLOSIS.CLIN.INF.DIS.14,131-140BROUQUI,P.ANDRAOULT,D.(2001)ENDOCARDITISDUETORAREANDFASTIDIOUSBACTERIA.CLIN.MICROBIOL.REV.14,177-207GADELRAB,M.O.ANDKAMBAL,A.M.(1998)EVALUATIONOFABRUCELLAENZYMEIMMUNOASSAYTEST(ELISA)INCOMPARISONWITHBACTERIOLOGICALCULTUREANDAGGLUTINATION.J.INFECT.36,197-201CORBEL,M.J.(1997)BRUCELLOSIS:ANOVERVIEW.EMERG.INFECT.DIS.3,213-221ZIMMERLI,S.UNDKÖPPEL,C.(2005)BRUCELLOSE.BVET-MAGAZIN3,36-37.AdditionalInformation:NameBrucellaIgGRelatedProductNamesBrucellaIgG–ELISAMolecularWeight0.5StorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°C.IntendedUseResearchUseOnly