Brucellosisisaninfectiousdiseasecausedbysmallellipsoid,gram-negativebacteria.Therearefourdifferentgerms:Br.abortus,Br.melitensis,Br.suisandBr.canis.Peopleareinfectedbycontactwithinfectedanimalsorbyeatingmeatordrinkingunpasteurizedmilkfrominfectedanimals.Asarule,infectedhumansarenotcontagious.Brucellosisismostfrequentinyoungandmiddle-agedindividuals.Endangeredpersonsarebutchers,farmers,ownersofpets,veterinariesandtouristsinSoutherncountries.Theappearanceofbrucellosisshowsaprevalenceduringwinterandspring.|Theincubationperiodisbetweenoneandthreeweeks,butmaybeaslongastwomonths.Br.abortusandBr.melitensismaycauseBang’sDisease,orinrarecasesMaltaFever.Thefirstappearsoccasionallywithalowpathogenicityforman.TypicalsymptomsforBang’sDiseaseareperiodicallyoccurringfever,splenomegalyandswellingoflymphnodes.Insomecasesaninflammationofdifferentjointsandorgansoccurs.TheMaltaFeveriscausedbytheepidemictypeofbrucellosis,andinfectionalmostalwaysleadstoamanifestillness.SomeinfectionswithBrucellacancauseBrucellaHepatitis.ItispossIBLethatthereisalinkbetweenaninfectionwithBrucellaandtheoutbreakofmultiplesclerosis.|Duringanantibiotictherapyorachronicinfection,thedetectionofBrucellaspec.inblood,urine,cerebrospinalfluid,sputumorotherbodyfluidscouldbenegative.SEROlogicalmethodslikeagglutination,complementfixationreaction,BrucellaCoombstestandELISAaregoodalternatives.Themonitoringofantibodiescanserveasausualindicationofthestatusofinfection.Duringthefirstdays,IgMistheonlyimmunoglobulinthatappears.Asthediseaseprogresses,IgMrecedesquantitativelyandIgGbecomespredominant.Inchronicbrucellosis,IgGmaybeproducedforanextendedperiod.
IntendedUse:|TheBrucellaIgGAntibodyELISATestKithasbeendesignedforthethedetectionandthequantitativedeterminationofspecificIgGantibodiesagainstBrucellainserumandplasma.FurtherapplicationsinotherbodyfluidsarepossibleandcanberequestedfromtheTechnicalServiceofUnitedStates
BIOLOGical.Thisassayisintendedforin-vitrodiagnosticuseonly.|Laboratoryresultscanneverbetheonlybaseofamedicalreport.Thepatienthistoryandfurthertestshaveadditionallytobetakenintoaccount.
PrincipleoftheTest
TheBrucellaIgGantibodytestkitisbasedontheprincipleoftheenzymeimmunoassay(EIA).Brucellaantigenisboundonthesurfaceofthemicrotiterstrips.Dilutedpatientserumorready-to-usestandardsare
Pipettedintothewellsofthemicrotiterplate.AbindingbetweentheIgGantibodiesoftheserumandtheimmobilizedBrucellaantigentakesplace.Afteraonehourincubationatroomtemperature,theplateisrinsedwithdilutedwashsolution,inordertoremoveunboundmaterial.Thenready-to-useanti-human-IgGperoxidaseconjugateisaddedandincubatedfor30minutes.Afterafurtherwashingstep,thesubstrate(TMB)solutionispipettedandincubatedfor20minutes,inducingthedevelopmentofabluedyeinthewells.Thecolordevelopmentisterminatedbytheadditionofastopsolution,whichchangesthecolorfrombluetoyellow.Theresultingdyeismeasuredspectrophotometricallyatthewavelengthof450nm.TheconcentrationoftheIgGantibodiesisdirectlyproportionaltotheintensityofthecolor.
KitComponents
Brucellaantigencoatedmicrotiterstrips,1x12stripsCalibratorA(NegativeControl),1x2mlCalibratorB(Cut-OffStandard),1x2mlCalibratorC(WeakPositiveControl),1x2mlCalibratorD(PositiveControl),1x2mlEnzymeConjugate,15mlSubstrate,1x15mlStopSolution,1x15mlSampleDiluent,1x60mlWashBuffer(10×),1x60ml
Storeallcomponentsat4°C.Stablefor6months.Reconstitutedstandardsarestablefortwoweeksstoredat4ºC.Formaximumrecoveryofproduct,centrifugetheoriginalvialpriortoremovingthecap.
References
1.DieterichRA,MortonJK,ZarnkeRL.J.Wildl.Dis.,27:470(1991).2.GoldsteinEJ.Infect.Dis.Clin.NorthAm.,5:117(1991).3.Greiser-WilkeI,MacMillanAP,MoennigV.Tierärztl.Prax.,19:131(1991).4.MurrellIG,MatthewsBJ.Med.Hypotheses,33:43(1990).5.SmithMC.Vet.Clin.NorthAm.FoodAnim.Pract.,6:705(1990).6.SuterO.Schweiz.Rundsch.Med.Prax.,80:1229(1991).7.WilliamsJD,HeckFC,DavisDSetal.VetImmunol.Immunopathol.,29:79(1991).8.YoungEJ.Rev.Infect.Dis.,13:359(1991).
