
Description:
AntibodiestoScl-70areaspecificimmunologicalMarkerforscleroderma(orprogressivesystemicsclerosis,PSS),asystemicautoimmunediseasecharacterizedbycollagendepositionandconnectivetissuedestructionoftheskin,bloodvesselsandcertaininternalorgans.StudieshaveshownvaryingfrequenciesofScl-70antibodiesinPSS.Thisantibodywasfoundinapproximately20%ofPSSpatientsinearlystudiesbut75%inlaterstudies.Scl-70antibodiesaredirectedagainstDNA-topoisomeraseIwhichlocatesinthenucleus.ThewholemoleculeofDNA-topoisomeraseis110kDabutitiseasilydegradedbyproteasesto100kDa,87kDaand70kDa(Scl-70).PSSisclassifiedintotwotypes;diffusesclerodermaandlimitedscleroderma.Scl-70antibodiesarepresentspecificallyindiffusesclerodermaandcentromereantibodiesarepresentinlimitedscleroderma.Rarely,Scl-70antibodiesarefoundinSLEandMCTDpatients.Principle:
AutoimmuneELISAkitsmeasureautoimmuneantibodiesintheserum.Itisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesaspecificantigenforimmobilizationonthemicrotiterwellsandanti-humanIgGantibodyconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwocomponents,resultingin autoimmuneantibodiesbeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofautoimmuneantibodies isdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm.