UtilizationGuide

VSpro

VSproPT/aPTTCombinationTest

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

TheVETSCANVSpromakespoint-of-carecoagulationtestingeasy,fast,andaffordable.ImmediatePTandaPTTtestresultsoffernumerousclinicalbenefitsforveterinarypatientsandfinancialbenefitsfortheveterinarypractice.

TheVETSCANVSproPTandaPTTCombinationTestisusedinconjunctionwiththeVSproAnalyzerforfast,convenient,andcost-effectivepoint-of-care coagulationtesting.ThistestcartridgeprovidesquantitativedeterminationofPTandaPTTsimultaneouslyfrom100µLofcitratedcanineorfelinewholeblood.

Cartridge Storage:Refrigerateat2°Cto8°C(39°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):1

Feline2

15-21

86-137

Canine1

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.3,4

• Hemolyzedsamplescanleadtotestingerrorsandinconclusiveresults.

•Thesodiumcitratetubemustbefilledcompletelyforaccurateresults.5

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 PTsecondsaPTT

seconds

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.

1

2

3 1 2 3

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6

4 5 6

7 8

9 10

7

8

9

10

VSpro

NormalCoagulationCascade

Likely Common Pathway or Combined Pathway Disorder:• Rodenticide toxicity• Vitamin K defi ciency• Factor XII• Hepatic disease • Bile insuffi ciency• DIC• Anticoagulants• Iatrogenic• Mast cell tumor

degranulation

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

Normal Prolonged

Normal NormalProlonged

Likely Extrinsic Pathway Disorder:• Factor VII• Early rodenticide

toxicity• Vitamin K defi ciency• Hepatic disease• Bile insuffi ciency

Prolonged

Ifbruisingorpetechia,consider:•VonWillebrand’sdisease•Thrombocytopenia

orplateletdisorder•Vasculardisorder

PT

aPTT aPTT

Test Interpretation6

PT (Prothrombin Time):MeasurestheExtrinsicandCommonPathways

aPTT (Activated Partial Thromboplastin Time): MeasurestheIntrinsicandCommonPathways

Suggested Test Utilization

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

HemophiliaA,orfactorVIIIdeficiency,isthemostcommoninheritedcoagulopathyofanimals.HemophiliaAismostprevalentinGermanSheparddogs.HemophiliaB,factorIX,deficiencyaffectscatsanddogs.Otherlesscommonhereditarycoagulationdeficiencieshavebeenreportedinanimalsaswell.7

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

Hepaticdisease8

Anypatientwithincreasedliverenzymes,possiblehepaticdysfunction,orconfirmedhepatopathywillbenefitfromcoagulationtesting.Thisbecomesimperativeshouldthepatientrequireinvasivesurgeryorbiopsy/aspirateofaninternalorgan.

Liverdiseasecanaffectthecoagulationcascadeinmultipleways,astheliverproducesmostofthecoagulationfactors.Considerthat:•Manyoftheclottingfactorsaresynthesizedandclearedbytheliver.•VitaminKisfatsoluble,soitsabsorptiondependsonadequatebileproductionandflow.

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

VitaminKdeficiencyorantagonismVitaminKisanessentialcofactorforcoagulationfactorsII,VII,IXandX.FactorVIIhastheshortesthalf-lifeandwilldepletetheearliest,therefore,PTisoftenprolongedfirst.9

SomecausesofVitaminKdeficiencyare:10

•Rodenticidetoxicity•Cholestaticliverdisease(reducedbileflowreducesabsorption)•LiverFailure•Malabsorptiondisorders•Medications

Otherdiseasestateswherecoagulationtestingisindicated:•Anypatientwithunexplainedbleeding,bruisingorpetechialhemorrhage•Snakebite •Infectiousdisease •Immune-mediateddisease•Shockorseveresystemicdisease;potentialforDIC(disseminatedintravascularcoagulopathy)•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:11, 12 Dynamic Range:11, 12

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

1

2

3

4

1 2

3 4

Test Procedure

VSpro

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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2

3 1 2 3

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5

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4 5

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Suggested Test Utilization

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

SystemicinflammatorymarkerEarlyrecognitionofsystemicinflammationusingfibrinogenisessentialforformulatinganeffectivetreatmentplan.Inflammationthatgoesunrecognizedcanimpairapatient’sgrowthandperformance.•Fibrinogenbloodlevelsaredirectlyrelatedtotheseverityoftheunderlyingcondition.•Fibrinogenisthepreferredinflammatorymarkerforearlydetection(subclinical)ofRhodococcusequipneumonia.13

•FibrinogenincreasesmorequicklyinsynovialfluidthanserumamyloidA(SAA)(byseveralhours)followingendotoxinadministrationintothejoint.14

•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.

Plasma Protein:Fibrinogen (PP:F) Ratio

Usefulintheinterpretationoffibrinogenconcentrationwhenhyperproteinemiaispresent.Thisratioiscalculatedas:

PP:FRatio=fibrinogenmg/dl

(plasmaTPmg/dl-fibrinogenmg/dl)

10-15Normal

>15Dehydration

1Dataonfile.StudyNo.TI-03294,ZoetisInc.2Dataonfile.StudyNo.TI-04203,ZoetisInc.3LippiG,PlebaniM,FavaloroE.Interferenceincoagulationtesting:focusonspurioushemolysis,icterus,andlipemia.SeminarsInThrombosisAndHemostasis.2013Apr;39(3):258-266.4LippiG,SalvagnoGL,MontagnanaM,Lima-OliveiraG,GuidiGC,FavaloroE.QualityStandardsforSampleCollectioninCoagulationTesting.SeminarsInThrombosisAndHemostasis.2012;38:565-575.5MontiP,ArcherJ.Qualityassuranceandinterpretationoflaboratorydata[Chapter2].BSAVAManualofCanineandFelineClinicalPathology.2016:12.6TopperMJ,WellesEG.Hemostatis[Chapter4].DuncanandPrasse’sVeterinaryLaboratoryMedicine:ClinicalPathology.2003:99-135.7TopperMJ,WellesEG.Hemostasis[Chapter4].DuncanandPrasse’sVeterinaryLaboratoryMedicine:ClinicalPathology,FifthEdition.2011:127.8BainPJ.Liver[Chapter7].DuncanandPrasse’sVeterinaryLaboratoryMedicine:ClinicalPathology,FifthEdition.2011:224.9TopperMJ,WellesEG.Hemostasis[Chapter4].DuncanandPrasse’sVeterinaryLaboratoryMedicine:ClinicalPathology,FifthEdition.2011:136.10Stokol,T.Disordersofhaemostasis[Chapter6].BSAVAManualofCanineandFelineClinicalPathology.2016:116.11Dataonfile.StudyNo.TI-03300,ZoetisInc.12EpsteinKL,BrainardBM.AnevaluationoftheAbaxisVSProforthemeasurementofequineplasmafibrinogenconcentrations.Equinevet.J.2012;44(4):449-452.13Passamonti,Fetal.Rhodococcusequipneumoniainfoals:anassessmentoftheearlydiagnosticvalueofserumamyloidAandplasmafibrinogenconcentrationsinequineclinicalpractice.VetJ.2015Feb;203(2):211-8.14Andreassen,Metal.Changesinconcentrationsofhaemostaticandinflammatorybiomarkersinsynovialfluidafterintra-articularinjectionoflipopolysaccharideinhorses.BMCVetRes.2017Jun19;13(1):182.15RobinsonNE,SprayberryK.CurrentTherapyinEquineMedicine,SixthEdition.2008:957.

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