VSproUtilizationGuide

VSproPT/aPTTCombinationTest

Coagulationtestingincludestheevaluationofbothprothrombintime(PT)andActivatedPartialThromboplastinTime(aPTT).Testingdeterminesifasignificantcoagulationfactordeficiencyexists,andifso,whichfactor(s)areaffected.PTisusedtoevaluatetheextrinsicandcommonpathways,whileaPTTisusedtoevaluatetheintrinsicandcommonpathways.

TheVetScanVSpromakespoint-of-carecoagulationtestingeasy,fastandaffordable.ImmediatePTandaPTTtestresultsoffernumerousclinicalbenefitsforveterinarypatientsandfinancialbenefitsfortheveterinarypractice.

TheVetScanVSproPTandaPTTCombinationTestisusedinconjunctionwiththeVSproAnalyzerforfast,convenientandcost-effective,point-of-carecoagulationtesting.ThistestcartridgeprovidesquantitativedeterminationofPT(prothrombintime)andaPTT(activatedpartialthromboplastintime)simultaneouslyfrom100µLofcitratedcanineorfelinewholeblood.

Cartridge Storage:Refrigerateat2°Cto8°C(36°Fto46°F)

Cartridge Stability: Donotexposecartridgesinthefoilpouchtodirectsunlight,anddonotleavethem atroomtemperature(15-30°C,59-86°F)formorethan3hours.

Usecartridgeswithin10minutesofremovingfrompouch.

Quick Reference Guide

Test/Species

PT

aPTTaPTT

Reference Ranges (seconds): Dynamic Ranges (seconds):

Feline

15-21

86-137

Canine

14-19

75-105

11-35

30-200

Sample well for adding 100 µL citrated whole

blood

Opticaldetectionwindows

Microfluidictechnology

Sample Preparation

Precautions before blood sample collection:

• Aswithanybloodtest,theaccuracyisdependentonthequalityofthebloodsample.Poorsamplequalityiscausedbyimpropercollectionandhandlingtechniques.

• Collectblooddirectlyfromthelargestaccessiblevein.

•Contaminationfromthromboplastin,alcohol,andintravenoussolutionswillinterferewiththecoagulationassay.

• Hemolyzedsamplescanleadtotestingerrorsandinconclusiveresults.

•Thesodiumcitratetubemustbefilledcompletelyforaccurateresults.

Fillanevacuatedsodiumcitratetubetothe“fill line”withwholeblood.Thebloodisdrawnintothetubeuntiltheflowstops.Forbloodcollectedintoaregularsyringe,theneedleshouldberemovedpriortogentlydispensingbloodintothetube.Ifnofilllineisseen,filltubetotopoflabel.NOTE:Under-filled or over-filled citrate tubes may alter results due to the improper anticoagulant to sample ratio, most commonly resulting in a falsely prolonged aPTT.

Fillline

1

2 Gentlyinvertthesodiumcitratetubeatleast10timesimmediatelyafterfillingtoensureagoodmixturewiththeanticoagulant.Letthesamplesitfor5minutesbeforetestingtostabilizethemixtureofbloodandcitrate.Ifthetestisdelayed,continuetoinvertevery10minutespriortothetest.Abloodrockermaybeused.NOTE:Blood may be tested up to two hours after it has been properly collected without affecting the test result.

Test Procedure

PT: seconds

aPTT: seconds

740-7021 Rev. B

Normal Range:

Species PTseconds

aPTTseconds

Dog 14 - 19 75 - 105

Cat 15 - 21 86 - 137

ResultsSticker

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RemovethePTandaPTTCombinationTestCartridgefromtherefrigeratorandopenthepouch.Touchthe“Analyze”buttonintheHomemenuscreentostartthetest.

Insertanewtestcartridgewhenthemessage “Please insert new cartridge”isdisplayedonthescreen.

Enterthe9-digit cartridge code locatedonthepouchlabelandtouch“Done”.

Selectspeciesandpress“Done”.

Touch“Confi rm”toacknowledgethatthesamplehasbeenobtainedinasodiumcitrate(bluetop)tube.

Enter“Patient ID”and/or“Sample ID”(optional).Touch“Done”toconfirmandthen“Next”tocontinue.The“Cartridge Warming Up”periodwillbegin.NOTE: Do not add sample while cartridge is warming up!

Whentheanalyzerbeepsandflashesthemessage“Add sample and wait”,usethedisposablepipetteprovidedtoaddsampletothewell.NOTE: Prior to adding sample, gently invert sample tube 10X to obtain a uniform mixture for testing.

Nextscreenshows“Test in progress”.Itlastsapproximately10minutesfromthetimethesampleisputintothewell.

Whenanalysisiscomplete,“Test Results”willappear.Usea“Results Sticker”(providedintheVSproStarterKitandcanbere-ordered:PN740-7021)toputthetestresultsintoafile;printresultsifprinterisattached;orsaveresultstoacomputerthroughproperconnectivity.(SeetheVSpromanualformoreinformation).TheVSprostoresupto1000testresults.Press“Done”toexitthetestresult.

“Please remove cartridge”thathasbeenusedanddisposeofitandalltestmaterialappropriately,andplacethesealingcartridgeinthecartridgeslot.

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NormalCoagulationCascade

LikelyCommonPathwayorCombinedPathwayDisorder:•Rodenticidetoxicity•VitaminKdeficiency•FactorXII•Hepaticdisease•Bileinsufficiency•DIC•Anticoagulants•Iatrogenic•Mastcelltumordegranulation

LikelyIntrinsicPathwayDisorder:•FactorVIII (HemophiliaA)•FactorIX (HemophiliaB)•FactorXI (HemophiliaC)•FactorXII•ExcessiveConsumption

Normal Prolonged

Normal NormalProlonged

LikelyExtrinsicPathwayDisorder:•FactorVII•Earlyrodenticide

toxicity•VitaminKdeficiency•Hepaticdisease•Bileinsufficiency

Prolonged

Ifbruisingorpetechia,consider:•VonWillebrand’sdisease•Thrombocytopenia

orplateletdisorder•Vasculardisorder

PT

aPTT aPTT

Test Interpretation

PT (Prothrombin Time):MeasurestheExtrinsicandCommonPathways

aPTT (Activated Partial Thromboplastin Time): MeasurestheIntrinsicandCommonPathways

Test Utilization

Pre-surgicaltestingPre-surgicaltestingshouldbeconsideredforanypatient,regardlessofage.Inheritedorcongenitalhemophiliacannotbedeterminedthroughanyothertestingmethodsoronphysicalexamination.Manyofthesedeficienciescausemildprolongationofclottingtimeswithoutclinicalsigns.

HemophiliaAandBhavebeendescribedinmanydogbreeds,includingmixedbreeds.HemophiliaAhasbeendetectedincats.HemophiliaCismorecommonincatsthanAorB.PT/aPTTtestingisanexcellentscreenfortoxiningestions(oftenlow-grade,subclinical)thatmaycauseprolongedbleedingtimes.Thisisfrequentlydetectedduringapre-surgicalcoagulationscreen.

PreventivecareBaselinevaluesareasimportantforcoagulationtestingastheyareforanyotheranalyte.Stress,illness,injury,medicationsandsurgerycanallaffectcoagulationtestresults,sobaselinevaluesarevitalforinterpretation.

HepaticdiseaseAnypatientwithincreasedliverenzymes,possiblehepaticdysfunction,orconfirmedhepatopathywillbenefitfromcoagulationtesting.Thisbecomesimperativeshouldthepatientrequireinvasivesurgeryorbiopsy/aspirateofaninternalorgan.

Liverdiseasecanaffectthecoagulationcascadeinmultipleways,astheliverproducesmostofthecoagulationfactors.Considerthat:•Manyoftheclottingfactorsaresynthesizedandclearedbytheliver.•VitaminKisfatsoluble,soitsabsorptiondependsonadequatebileproductionandflow.•Severeliverdiseasemayleadtodisseminatedintravascularcoagulation(DIC).

Anydiseasestatethataffectsthelivercanleadtoacoagulationabnormalityincluding:•Inflammation(hepatitis,cholangiohepatitis)•Neoplasia•Biliarystasis•Useofchronicmedications(NSAIDs,anesthetics,chemotherapeutics,etc.)

VitaminKdeficiencyorantagonismVitaminKisanessentialcofactorforcoagulationfactorsII,VII,IXandX.FactorVIIhastheshortesthalf-lifeandwilldepletetheearliest,therefore,PTisoftenprolongedfirst.SomecausesofVitaminKdeficiencyare:•Rodenticidetoxicity•Cholestaticliverdisease(reducedbileflowreducesabsorption)•LiverFailure•Malabsorptiondisorders•Medications

Otherdiseasestateswherecoagulationtestingisindicated:•Anypatientwithunexplainedbleeding,bruisingorpetechialhemorrhage•Snakebite •Infectiousdisease •Immune-mediateddisease•Shockorseveresystemicdisease(potentialforDIC)•Activelybleedingpatients

VSproFibrinogenTest

Sample well for adding diluted platelet-poor plasma

Opticaldetectionwindows

Microfluidictechnology

Quick Reference Guide

TheVetScanVSproFibrinogenTestcartridgeisusedinconjunctionwiththeVSproAnalyzertoevaluatethefibrinogenlevelinhorses,asensitiveandspecificmarkerforinflammation.Earlyrecognitionofsystemicinflammationisessentialfordiagnosisofdiseaseandformulatingatreatmentplan.Quantitativeserialtestingforfibrinogencanhelpeffectivelymonitortreatmentandtheresponsetotherapy.

Cartridge Storage:Refrigerateat2°Cto8°C(36°Fto46°F)

Cartridge Stability: Donotexposecartridgesinthefoilpouchtodirectsunlight,anddonotleavethem atroomtemperature(15-30°C,59-86°F)formorethan3hours.

Usecartridgeswithin10minutesofremovingfrompouch.

Equine

Reference Range: Dynamic Range:

mg/dL

150-400

g/L

1.5-4.0

g/L

0-20

mg/dL

0-2000

Sample Preparation

Precautions before blood sample collection:

• Aswithanybloodtest,theaccuracyisdependentonthequalityofthebloodsample.Poorsamplequalitycanbecausedbyimpropercollectionandhandlingtechniques.

• Collectblooddirectlyfromthelargestaccessiblevein.

•Contaminationfromthromboplastin,alcoholandintravenoussolutionswillinterferewiththefibrinogenassay.

• Useofhemolyzedsamplesmayresultinlowerthanexpectedvalues.

•Ensureprecisionwhenmixingplatelet-poorplasmaanddiluent,andonlyusethepre-filledmicrotubesupplied.

•Theplatelet-poorplasmaanddiluentsolutionmustbemixedgently.Donotuseavortexmixer,anddonotshakethesample

•Thetubemustbefilledcompletelyforaccurateresults.

Fillanevacuatedsodiumcitratetubetothe“fill line”withwholeblood.Thebloodisdrawnintothetubeuntiltheflowstops.Forbloodcollectedintoaregularsyringe,theneedleshouldberemovedpriortoplacingbloodintothesodiumcitratetube.Donotplacesampleonarocker.NOTE:Under-filled or over-filled citrate tubes may alter results due to the improper anticoagulant-to-sample ratio.

Spinthecitratedwholebloodsampletoplatelet-poorplasmausingacentrifuge.Refertopackageinsertforcentrifugerequirements.

Useapipetteandoneofthesuppliedpipettetipstoextract100µLofplatelet-poorplasmafromthecentrifugedsampletube.Depositthe100µLofplatelet-poorplasmaintothepre-filleddiluenttubebypiercingthefoillidwiththepipettetip.

Gentlymixthediluentandsamplebyextractinganddepositingthemixtureusingthepipette.Thesampleisnowreadyfortesting.

Fillline

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2

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4

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3 4

Test Procedure

ResultsSticker

RemovetheFibrinogenTestCartridgefromrefrigeration10-15minutespriortoanalysistowarmuptoroomtemperature.Touchthe“Analyze”buttonintheHomemenuscreentostartthetest.

Insertanewtestcartridgewhenthemessage“Please insert new cartridge”isdisplayedonthescreen.

Enterthe9-digitcartridge codelocatedonthepouchlabelandtouch“Done”.

Touch“Confirm”toacknowledgethatthesamplehasbeenobtainedinasodiumcitratetube,spundowntoplatelet-poorplasmaanddilutedbyadding100µLoftheplasmatothesuppliedpre-filleddiluenttube.

Enter“Patient ID”and/or“Sample ID”.Touch“Done”toconfirmandthen“Next”tocontinue.The“Cartridge Warming Up”periodwillbegin.NOTE:Do not add sample while cartridge is warming up!

Whentheanalyzerbeepsandflashesthemessage“Add 100 µL sample and wait”,usepipetteanddisposablepipettetiptoadd100µLofsampletothewell.NOTE:Prior to adding sample, extract and re-deposit the sample in the tube to obtain a uniform mixture of plasma and diluent for testing. Do not introduce bubbles to the mixture.

Nextscreenshows“Test in progress”.Itlastsapproximately15minutesfromthetimethesampleisputintothewell.

Whenanalysisiscomplete,“Test Results”willappear.Usea“Results Sticker”(providedintheVSproStarterKitandcanbere-ordered:PN740-7041)toputtestresultsintoafile;printresultsifprinterisattached;orsaveresultstoacomputerthroughproperconnectivity.(SeetheVSpromanualformoreinformation).TheVSproautomaticallystoresupto1000testresults.Press“Done”toexitthetestresult

“Please remove cartridge”thathasbeenusedanddisposeofitandalltestmaterialappropriately,andplacethesealingcartridgeinthecartridgeslot.

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Test Utilization

Fibrinogentestingside-by-sidewithcompletebloodcount(CBC)FibrinogenconcentrationsshouldbeincludedwhenperformingaCBCinthehorse.•Fibrinogenisanacutephaseproteinmadeintheliverthatdetectsinflammation.•Fibrinogenincreases24hoursafteraninflammatoryprocesshasstartedandpeaksbetween4and7days.•Fibrinogenmayindicateinflammationinthesubclinicalhorsewhentheleukogramiswithinnormallimits.•Fibrinogenlevelsareverysensitive,andevenamildincreaseshouldbeconsideredsignificant.

SystemicinflammatorymarkerEarlyrecognitionofsystemicinflammationusingfibrinogenisessentialforformulatinganeffectivetreatmentplan.Inflammationthatgoesunrecognizedcanimpairapatient’sgrowthandperformance.•Fibrinogenbloodlevelsaredirectlyrelatedtotheseverityoftheunderlyingcondition.•Fibrinogenisthepreferredinflammatorymarkerforearlydetection(subclinical)ofRhodococcusequipneumonia.1

•FibrinogenincreasesmorequicklyinsynovialfluidthanserumamyloidA(SAA)(byseveralhours)followingendotoxinadministrationintothejoint.2

•Whentestedserially,fibrinogenprovidesinformationregardingtreatmentefficacyandlength,prognosis,especiallyconsideringsuchinfectiousorinflammatoryconditionsas:R.equiinfections,strangles,pigeonfever,pleuropneumonia,abdominalabscess,septicarthritis,endometritis,clostridialmyonecrosis,andendocarditis.

Fibrinogenspecificity•Plasmafibrinogenmayincreasewithdehydrationandmaydecreasewithseverehepaticdiseaseduetodecreasedproteinproduction.•IncasesofDIC,fibrinogenmaybedecreasedduetoincreasedutilization(maskingthehyperfibrinogenemiaassociatedwiththeinflammatoryprocess).

Patientmonitoringtool•Thedegreeofhyperfibrinogenemiaapproximatestheseverityofdisease.•Whenused,inconjunctionwiththeclinicalfindingsandotherlaboratorydata,themosteffectivetreatmentprotocolandprognosiscanbemonitoredclosely.•Fibrinogenlevelsmaybeusedaloneindeterminingresolutionoftheinflammatoryprocesswhentheleukogramisunchangedorhasreturnedtonormal.

1.Passamonti,Fetal.Rhodococcusequipneumoniainfoals:anassessmentoftheearlydiagnosticvalueofserumamyloidAandplasmafibrinogenconcentrationsinequineclinicalpractice.VetJ.2015Feb;203(2):211-8.2.Andreassen,Metal.Changesinconcentrationsofhaemostaticandinflammatorybiomarkersinsynovialfluidafterintra-articularinjectionoflipopolysaccharideinhorses.BMCVetRes.2017Jun19;13(1):182

Plasma Protein:Fibrinogen (PP:F) Ratio

Usefulintheinterpretationoffibrinogenconcentrationwhenhyperproteinemiaispresent.Thisratioiscalculatedas:

PP:FRatio=fibrinogenmg/dl

(plasmaTPmg/dl-fibrinogenmg/dl)

15-20Normal

>20Dehydration

<15Inflammation

Interpretation:

TheVetScanVSproisintendedforveterinaryuseonly.AbaxisandVetScanareregisteredtrademarksofAbaxis,Inc.©Abaxis2018

GLOBAL DIAGNOSTICS

888-2002Rev.D

Abaxis, Inc.

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