Description:
REAADSAnti-ProthrombinIgG isan anti-ProthrombinantibodyELISAtestkitforthe semi-quantitative,indirectdetectionofantibodiesagainsthumanprothrombin.AsinglepointcalibratorisusedtodetermineIgGanti-prothrombinantibodyconcentrationsinhumanserumofcitratedplasma.
Advantages:
- ConvenientELISAprocedure
- ForthespecificdeterminationofIgGisotype
- Objective,accurateandreproducIBLe
- Reagentcompletekits
- Totalincubationtime:40minutesatroomtemperature
PrincipleandProcedure:
Principle
TheREAADSIgGanti-Prothrombinantibodytestkitsaresemiquantitative,indirectELISAsforthedetectionofantibodiesagainsthumanprothrombin.AsinglepointcalibratorisusedtodetermineIgGanti-prothrombinantibodyconcentrationsinhumanserumofcitratedplasma.
Procedure
Dilutedpatientserumorplasmaisincubatedinmicrowellscoatedwithhumanprothrombin.Anti-Prothrombin(aPT)antibodiesinpatientsampleswillbindtotheimmobilizedantigeninthewells.Afterwashingtoremoveunboundserumorplasmaproteins,antibodiesspecificforhumanIgG,labeledwithhorserADIshperoxidase(HRP),areboundantibodies.Thewellsarewashed,andachromogenicsubstrateisadded,resultinginacolorchangethatisproportionaltotheamountofantibodypresent.Theassayiscompletedbytheadditionofastoppingsolution.ResultsareobtainedbyreadingtheO.D.(opticaldensityorabsorbance)at450nmofeachwellinaspectrophotometer.TheO.D.valuesofcontrolandpatientsamplesaremultipliedbythecalibratorconversionfactortoobtainIgGaPTantibodyconcentrationstimeis40minutesatroomtemperature.
ClinicalPerformance*:
ClinicalSpecificity:serumsamplesfrom100healthyblooddonorsweretestedforaPTantibodies.Againstanestablishedcutoffvalueof20Gand20Munits,thespecificityoftheIgGassaywas95%.Aspresentedinthetablebelow,similarresultswereseenwhenplasmasamplesfrom100healthyblooddonorsweretested.Assayspecificitywith42infectiousdisease(syphilis)patientsand42RheumatoidArthritis(RA)patientsisalsopresentedinthetable.
ClinicalSensitivity:serumsamplesfrom41unselectedSLEpatientsweretestedforIgGaPTantibodies.14.6%ofthesampleswerepositiveforIgGaPTantibodies(meanvaluesof10.4Gunits).Serumsamplesfrom11selectedfemalepatientswithprimaryanti-phospholipidsyndrome(APS)werealsoevaluatedforantibodiestoprothrombin.18.2%ofthesampleswerepositiveforIgGantibodies(meanvaluesof12.3G).
| IgGaPT (%positive) | |
| Healthyserum (n=100) | 5% |
| Healthyplasma (n=100) | 1% |
| Syphilis (n=42) | 4.8% |
| RA (n=42) | 4.8% |
| SLE (n=41) | 14.6% |
| APS (n=11) | 18.2% |
*Dataavailableuponrequest.
Background:
Severalplasmaproteinsassociatedwiththecoagulationsystemandwithstrongphospholipidbindingpropertieshavebeenidentifiedasantiphospholipidcofactors.Antibodiestoprothrombin,oneoftheseplasmaproteins,havebeenreportedinpatientswithantiphospholipidsyndrome.TestingfortheseantibodiesisproposedtobeincludedintheSEROlogicevaluationofantiphospholipidantibodies.Clinically,elevatedserumlevelsofanti-prothrombin(aPT)antibodiesareassociatedwithanincreasedriskforAPS,characterizedbyrecurrentarterialorvenousthrombosis,thrombocytopeniaand/orfetalloss.HighserumorplasmalevelsofaPTantibodiesmayaddvaluableinformationinthelaboratoryassessmentofantiphospholipidantibodies.
