LabEX2.docx
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LabEX2.docx
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LabEX2
LearningResources
1.BectonDickinson.(1992).TheBBL®FibroSystemManual,4thedition.BectonDickinsonMicrobiologySystems.Cockeyville,MD.Cat.No.98144.
2.Burns,Cheryl.LaboratoryTestinginCoagulation(Chapter39).In:
McKenzie,SB(ed.).ClinicalLaboratoryHematology(2004).UpperSaddleRiver,NJ:
PearsonPrenticeHall;2004.787–795.
3.UWDepartmentofLaboratoryMedicine,ReferenceLaboratoryServices,HandbookofDiagnosticHemostasisandThrombosisTests,3rdEdition,2005.6,7,m22,25–29,32,and33.Availableon-lineatURLhttp:
//depts.washington.edu/labweb/PatientCare/Clinical/Guides/hemostasis.pdf
4.LaboratoryTestsOn-lineatURLhttp:
//www.labtestsonline.org/
LearningObjectives–Uponcompletionofrequiredreading,aftercarefulstudy,andfollowingthislabexercise,thelearnerwillbeableto:
1.DescribethetestprincipleforthelatexslideagglutinationtestforD-dimers.
2.CorrectlyinterpretD-dimerslidetestresultsandcorrelatewithotherlabtestinformationandmedicalhistorytoaffirmitsclinicalutility.
3.PerformthePT,PTT,TT,andquantitativefibrinogenassaysusingthefibrometerandobtainsatisfactoryresultsthatcanbeaccuratelyinterpretedbycoursestudentsandinstructors.
4.Assessfibrinogenclottingtimesandusetheinformationasaguidetomakeappropriateplasmasampledilutionsincaseswhenthetimesareshortorlong.
5.Judgetheacceptabilityoftestresults,includingqualitycontroldata,andimplementappropriatecorrectiveactionswhenerrorsareapparent.
6.Evaluatecoagulationscreentestandmixingstudyresultscollectivelytonarrowadiagnosisforapatientandrecommendadditionalteststoconfirmdiagnoses.
7.Constructanduseastandardcurvetodeterminetheconcentrationoffibrinogeninclinicalplasmasamples.
8.Followwrittenandverbalinstructions,andprudentlymanagetimeandresourcestocompletethelabexerciseinthetimeallotted.
9.Demonstratemulti-taskingskillsusingautomationtoperformtestingusefulinevaluatingbleedingdisorders.
10.Predictcoagulationscreentestandmixingstudyresultsincasesof:
a)functionalfactordeficiency,b)immediate-actinginhibitors,andc)time/temperaturedependentinhibitors.
11.Differentiateareferencerangefromanacceptablerangeforcontrols.
LabReport:
Dueone
(1)weekfromtoday.Reportistoincludeatitlepage,pages4-6,and8–15.
YourLearningAssignments
1.Beforecomingtoclass,READforunderstandinglearningreferences2,3,thislabexercise,andtheProthrombintime(PT),PartialThromboplastintime(PTT),Thrombintime(TT),Fibrinogen(Fi),MixingStudy,andtheD-dimerprocedurespostedonthecoursewebpageatURL:
http:
//faculty.washington.edu/calvoc/HomePage/LabM419Labs.shtml).
2.Inclass,FIRSTset-upyourworkstation:
DIRECTIONSTOSET-UPYOURWORKSTATION
a)Checkthatyourfibrometersandheatblockunitareturnedonandtheicebucketisfilled.
b)Obtainandpreparereagents(i.e.,reconstitute,aliquot,placeiniceorheatblock,etc.)asmaybeneededandtestsamplestobeusedintheassaysyou’reperformingtoday.
3.Inclass,aftersettingupyourworkstation,ASKquestion(s)duringthebriefdiscussionRE:
mixingstudiesandduringtheshortdemonstrationontheslidelatexagglutinationtestforD-dimers.
4.Lastly,followtheprocedures(PT,PTT,TT,Fi,andmixingstudies)tocompletethetestingindicated.
a.RunPT,PTT,TT,FibrinogenontheNormalControl
b.RunPT,PTTontheAbnormalControl
c.RunPT,PTT,TTandFibrinogenonUnknownplasmas1and2,andperforma1:
1mixusingthenormalcontrolasthediluentplasmaforanytestforwhichitmaybeappropriate.EXTENDEDincubationat37°Cisnotrequiredforthisexercise.
d.RunafibrinogenonUnknownplasma3andanyfollow-uptestingnecessaryinordertoobtainandreportatruefibrinogenresultforthissample.
NOTE:
YouwillNOTbeperformingthetestingnecessarytoconstructastandardcurvethatcanbeusedtodeterminetestplasmafibrinogenconcentrations!
!
!
!
!
!
!
!
!
Astandardcurvewillbeprovidedforyou.
5.BesuretodiscussyourQCandfibrinogenclottimeresultswithaninstructortoverifyyourunderstanding.
YourLearningAssignments–continued
StandardLaboratoryPractices
1.Beginningwiththislabexercise,andcontinuingwithallremaininglabexercises,youareexpectedtoevaluatequalitycontrol(s)dataandduplicateresults,andtakeappropriatecorrectiveactionsasneededBEFOREproceedingwithfurthertesting(i.e.,unknowns,etc.).Yourlabexercisedatawillbegradedaccordingly.IFaninstructoryouareworkingwithinclassdirectsyoutoproceedwithtestingdespitecontroldatabeing‘out’,THENyoumusthavethatinstructorinitialanddateyourdatasheetasprooforpointswillbedeductedforinaccuracyandfailuretofollowstandardlaboratorypractice.
2.Mixreagentaliquotswellpriortopipettingthemfortestinginordertobreakupaggregatesthatmayformovertimeasthesolution(s)incubateatvarioustemperatures.
3.Iftestingmustberepeated,recordinitialresults,andrepeatresults.Alwayscircletheresult(s)thataretobereported.
4.Forthiscourse,thenormalcontrolisusedastheplasmadiluentwhenconductingamixingstudy.Somelabmayusepooledplasmafromhealthyadultdonors.However,beforepoolednormalplasmaisusedasadiluent,itistestedbyperformingtheassay(s)whichareusedwhenconductingamixingstudy(e.g.,PTTorTT).Thisisdonetoverifythatthepooledplasmais‘normal’.Clinicallaboratoriesmayalsochoosetopurchase(e.g.,CRYOcheck™PooledNormalPlasma,PrecisionBioLogic,Dartmouth,NovaScotia)theyneedformixingstudies.
PARTA–D-dimerDemonstration
Tocompletethisactivityyouneed:
∙Thegraphiconwhichtorecordyourobservations
∙Aninkpen
DIRECTIONS:
Instructorswilldemonstratethetest.Usethegraphicbelowtorecordwhatyouobserve.
(2)
StudyQuestions–Pointvaluesappearinparentheses.
1.WhatareD-dimers?
(1)
Exercise2PartA:
StudyQuestions–continued
2.BrieflydescribethetestprincipleforthelatexslideagglutinationtestforD-dimers.
(1)
3.DescribetheclinicalvalueoftheD-dimerassay.
(1)
PARTB–DifferentiatingReferenceRangesandAcceptableRangesforControls
Tocompletethisactivityyouneed:
∙Thechartstorecordinformation
∙Aninkpen
DIRECTIONS:
Fill-intherequestedinformationtocompletethechartsbelow.
ReferenceRanges(4)
TheTest
ReferenceRange
Prothrombintime
PartialThromboplastinTime
ThrombinTime
Fibrinogen
AcceptableRangesforControls(8)
TheTest
NormalControl
AbnormalControl
Prothrombintime
PartialThromboplastinTime
ThrombinTime
Fibrinogen
StudyQuestion-Pointvaluesappearinparentheses.
1.Brieflyexplainthedifferencebetweenthesetwodatasets.(HINT:
Whatarethesedatausedfor?
)
(1)
PARTC–EvaluatingBleedingDisorders
Tocompletethisactivityyouwillneed:
∙TheproceduresforthePT,PTT,TT,Fi(Fibrinogenquantitation)andmixingstudies
∙Datasheet
∙Reagents,controls,materials,andsuppliesspecifiedintheprocedures
∙Fibrometersoutfittedwitha0.3mLprobes
6.DIRECTIONS–Followtheprocedures(PT,PTT,TT,Fi,andmixingstudies)tocompletethetestingindicated.
e.RunPT,PTT,TT,FibrinogenontheNormalControl
f.RunPT,PTTontheAbnormalControl
g.RunPT,PTT,TTandFibrinogenonUnknownplasmas1and2,andperforma1:
1mixusingthenormalcontrolasthediluentplasmaforanytestforwhichitmaybeappropriate.EXTENDEDincubationat37°Cisnotrequiredforthisexercise.
h.RunafibrinogenonUnknownplasma3andanyfollow-uptestingnecessaryinordertoobtainandreportatruefibrinogenresultforthissample.
NOTE:
YouwillNOTbeperformingthetestingnecessarytoconstructastandardcurvethatcanbeusedtodeterminetestplasmafibrinogenconcentrations!
!
!
!
!
!
!
!
!
Astandardcurvewillbeprovidedforyou.
7.BesuretodiscussyourQCresultsandyourfibrinogenclottimedatawithaninstructortoverifyyourunderstanding.
LabExercise2PartCDATASHEET(172)
Reagent
Lot#
ExpirationDate
PT(STA®NeoplastineC1Plus)
PTT(STA®PTT)
CaCl2(STA®CaCl20.025M)
STA®Fibrinogen5
STA®Owren-Kollerbuffer
STA®Thrombinreagent
Cryo√PNP–Normalcontrol
Cryo√-Abnormal2control
Sample
PT
(s)
PTT
(s)
TT
(s)
Fi
(s)
Fi(mg/dL)
1
2
Ave
1
2
Ave
1
2
Ave
1
2
Ave
NCtrl
ABNCtrl
UNK1
(Screen)
UNK2
(Screen)
UNK3
(Fionly)
LabExercise2PartCDATASHEET-continued
StudyQuestions:
–Pointvaluesappearinparentheses.
1.Foreachunknown,brieflydiscussthevalidityofeachtestresult.(HINT:
Whatcriteriadidyouusetoevaluatethedatavalidity?
Iftheinitialresultswerenotreportable,whatdidyouhavetodonexttoobtainvalidresults?
Why?
)(10)
LabExercise2PartCStudyquestions-continued
2.Fortheunknownplasma(s)onwhichyouperformedcoagulationscreensand1:
1mix(es)whenappropriate,reviewthedata.
a.Whatdotheresultssuggestregardingtheetiologyoftheresults:
factordeficiencyorinhibitor?
(1)
b.Brieflydiscussthesignificantresultsthatledyoutothisconclusion.Ifyouconcludedafactordeficiency,expl
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