Description:
ChromogenixCoatest®APC™Resistance–VSisanAPTT-basedassaykitforthescreeningoffactor-V-relatedAPCresistance.ThehighsensitivityandspecificityofthetestforthefactorV:Q506mutationisobtainedbypredilutingthesampleplasmawithanexcessofV-DEFPlasmabioreagent.ThetestdesignmakesitpossIBLetodiscriminatebetweenheterozygousandhomozygousfactorVgenotypes.Italsoallowsforanalysisofplasmafrompatientsonheparinororalanticoagulanttherapy.Highdiscriminationbetweengenotypeswith100%sensitivityforFV:Q506.ReducesneedforPCRdetermination.Applicabletoanticoagulanttreatedpatients.
Atime-andmoney-savingalternativetoindividualgenetictesting,Coatest©APC™Resistancetestingisanidealsolutionforphysiciansandlabs.Eachkitwasdesignedtobeeasytouseforin-housescreening,offeringfastresultsandlowcostsforbothlarge-andsmall-volumelabs.ThemostcommonFVLeidenscreeningtestperformed,itoffersunmatchedsensitivityfortheFV:Q506mutation–closeto100%specificity–andisapplicabletopatientsonheparinorwarfarin.
MeasurementPrinciple:
OnevolumeofplasmaispredilutedwithfourvolumesofV-DEFPlasma.ThedilutionisthenincubatedwiththeAPTTreagentforastandardperiodoftime.CoagulationistriggeredbytheadditionofCaCl2intheabsenceandpresenceofexogenousAPCandthetimeforclotformationisrecorded.
ActivatedProteinC(APC)isaregulatorofthecoagulationcascade,byspecificallyinactivatingfactorsVaandVIIIa,inthepresenceofphospholipidsandcalcium. Inmostofthecases(morethan90%),ActivatedProteinCResistance(APCR)phenotypeiscausedbyaFactorVgenemutation(FactorVLeiden).Themutation,locatedonFactorVexon10(1691G–>A),ofargininetoglutamineonposition506,rendersFactorVaresistanttothecleavagebyActivatedProteinC.ThisgeneticanomalycanbeevidencedwithaclottingmethodperformedinthepresenceortheabsenceofActivatedProteinC.
KitComposition:
ReagentsandStABIlity:
1.V-DEFPlasma2vialsStabilized,lyophilizedhumanplasma,withalowleveloffactorVactivity,containingtheheparinantagoNISTPolybrene®.Reconstitutewith4.0mLofNCCLStypeIIwater.Allowtostandfor30minutesat20-25°C.Swirlgentlybeforeuse.
2.CaCl22vials2mLofcalciumchloride,0.025mol/L,inTrisbuffercontaining0.5%bovineserumalbumin.
3. APTTreagent2vials4mLofpurifiedphospholipidswithcolloidalsilicaascontactactivator.Containsapreservative.MixthoroughlyonaVortexmixerbeforeuse.
4.APC/CaCl22vialsHumanactivatedproteinCcolyophilizedwithCaCl2.Reconstitutewith2.0mLofNCCLStypeIIwater.Allowtostandfor30minutesat20-25°C.Swirlgentlybeforeuse.
5.ControlPlasmaLevel1.1vialLyophilizedhumanplasma.Reconstitutewith1.0mLofNCCLStypeIIwater.Allowtostandfor30minutesat20-25°C.Swirlgentlybeforeuse.
6.ControlPlasmaLevel2.1vialLyophilizedhumanplasma.Reconstitutewith1.0mLofNCCLStypeIIwater.Allowtostandfor30minutesat20-25°C.Swirlgentlybeforeuse.
TestingAlgorithm:
IDEALFOR:
- Labslookingtobringtestingin-house
- Labslookingforcostsavings
- Orderingphysicianslookingforasensitiveassaywithrapidturnaround
- Savesthelabfromhavingtoperformgenetictestonallpatients
- Economicalalternativetogenetictesting
Background:
ActivatedProteinC(APC)isaregulatorofthecoagulationcascade,byspecificallyinactivatingfactorsVaandVIIIa,inthepresenceofphospholipidsandcalcium.
Inmostofthecases(morethan90%),ActivatedProteinCResistance(APCR)phenotypeiscausedbyaFactorVgenemutation(FactorVLeiden).Themutation,locatedonFactorVexon10(1691G–>A),ofargininetoglutamineonposition506,rendersFactorVaresistanttothecleavagebyActivatedProteinC.ThisgeneticanomalycanbeevidencedwithaclottingmethodperformedinthepresenceortheabsenceofActivatedProteinC.
ActivatedProteinCResistanceistestedbyusingaclottingmethodperformedwithorwithoutActivatedProteinC.
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