Description:

Technothrombin® TGAisathrombingenerationassay(TGA)basedonmonitoringtheformationofthrombinbymeansofafluorogenicsubstrateuponactivationofthecoagulationcascadebytissuefactor.

TheuseofTechnothrombin® TGAforsuchdiverseapplicationsismadepossIBLebecauseTechnothrombin® TGAallowstomeasurethewholekineticsofthrombingenerationnotonlyduringtheinitiationphaseofthrombinformationwiththeendpointfibrinformation,butalsoduringthephaseofdownregulationofthrombinformationandinactivationoftheformedthrombin.Technothrombin® TGAisthereforeauniversaltoolforanalysisandresearch ofthehemostaticsystemonanindividualbasis.

KitComposition:

• Substrate:3x1.5TGAsubstrate(SUB)Fluorogenicsubstrate1mMZ-G-G-RAMC, 15mMCaCl2
• Buffer:1x3TGAbuffer(BUF)Hepes-NaCl-buffercontaining0.5% bovineserumalbumin
• Calibrator:1x0.5TGAthrombincalibrator~1.000nMthrombininbufferwithBSA
• ReagentCLow:1x0.5TGAreagentC(RC)LowRCLowconc.ofphospholipidmicelles containingrhTFinTris-Hepes-NaClbuffer
• ReagentCHigh:1x0.5TGAreagentC(RC)HighRCHighconc.ofphospholipidmicelles containingrhTFinTris-Hepes-NaClbuffer
• ReagentD:1x1.5TGAreagentD(RD)RDconc.ofphospholipidmicellesinTrisHepes-NaCl buffer
• ControlHigh:1x1TGAcontrolhigh(CH)Humanplasmawithincreasedthrombin generation,lyophilized.
• ControlLow:1x1TGAcontrollow(CL)Humanplasmawithdecreasedthrombin generation,lyophilized.

Eachreagentisavailableseparately.

MeasurementPrinciple:

Technothrombin® TGAisbasedonmonitoringthefluorescencegeneratedbythrombincleavageofafluorogenicsubstrateovertimeuponactivationofthecoagulationcascadebydifferentconcentrationsoftissuefactor(RA,RB,RCLowandRCHigh)andnegativelychargedphospholipidsinwholebloodorplasma.Fromthechangesinfluorescenceovertime,theconcentrationofthrombin(nM)inthesamplecanbecalculatedusingtherespectivethrombincalibrationcurve.Therateofincreaseinthrombinconcentrationovertimethenallowstocalculategenerationofthrombininthesampleperminuteandtoplotthisvalueovertimeforthewholecoagulationprocess.Thisthenresultsinthevisualizationofthedifferentphasesofclotformation.

Background:

TheTechnothrombin® TGAkitisanassaysystemfordeterminationof thrombingenerationovertimeinplateletpoororplateletrichplasma(PPPor PRP)uponactivationoftheclottingcascadebymicellesofnegativelycharged phospholipidscontainingdifferentamountsofhumantissuefactorandCaCl2.The kitcanbeusedtomonitorhemophiliacsduringinhibitorbypassingtherapy,to monitoranticoagulationtherapy,tocalculateINRvaluesforpatientsandto determinestatesofbleedingdisordersorthrombophiliaaswellastheactivityof circulatingmicroparticles.Thisbroadrangeofapplicationsispossiblebyproviding differenttissuefactorconcentrationsandbymonitoringthewholekineticof thrombingenerationduringinitiation,amplificationanddownregulationofthrombin formation.Technothrombin® TGAisthereforeauniversalassaykitfor analyzingandresearching thefunctionofthehemostaticsystem.