Compatibility-Testing：兼容性测试完整版资料

CompatibilityTestingAhmadShihadaSilmiMsc,FIBMSIUGWhatiscompatibilitytesting?AlsocalledpretransfusiontestingPurpose:ToselectbloodcomponentsthatwillnotcauseharmtotherecipientandwillhaveacceptablesurvivalwhentransfusedIfproperlyperformed,compatibilitytestswillconfirmABOcompatibilitybetweenthecomponentandtherecipientandwilldetectthemostclinicallysignificantunexpectedantibodiesCompatibilitytesting?ThereareseveralcomponentsofcompatibilitytestingProperspecimencollectionReviewingpatienttransfusionhistoryABO,Rh,andantibodytesting(screen/ID)CrossmatchingActualtransfusionCompatibilitytestingCanbedividedinto3categories:PreanalyticalproceduresSerologicaltestingPostanalyticalproceduresPre-analyticalphasesPatientidentificationSpecimencollectionReviewofpatienthistoryPatientIdentificationMustconfirmrecipient’sIDfrombraceletONthepatientFullpatientnameandhospitalnumberNameofphysicianSampleIdentificationThesampleshouldalsohavethefullpatientname,hospitalnumber,andphysicianDateandtimeofcollection,phlebotomist’sinitialsAllofthisshouldbeontherequestformandthesampleSpecimenTubesPinkTop-EDTARedTop–noadditivesSpecimenCollectionCollectedintubewithEDTAornoadditivesIfthevenipuncturecauseshemolysis,thesamplemayberejectedTruehemolysisinthepatientistheresultofcomplementactivationSamplesarelabeledatthebedside(pre-labelingisnotrecommended)Arecordofindividualswhocollect(ortest)thespecimensshouldbedocumentedinorderto“backtrack”incaseofanerrorSpecimenCollectionIfthesampleisdrawnfromanIVline,theIVinfusionshouldbestopped5-10minutespriortoblooddrawingandthefirst10mLdiscardedTestingshouldbeperformedonsampleslessthan72hoursorelsecomplementdependentantibodiesmaybemissed(complementcanbecomeunstable)GettingthehistoryLookatrecipient’srecordsforanypriorunexpectedantibodiesPrevioustransfusionreactionsSerologicalTesting3tests:ABO/RhAntibodydetection/identificationCrossmatchABO/RhTypingIntheABOtyping,theforwardandreverseMUSTmatchIntheRhtyping,thecontrolmustbenegativeBothofthesewillindicatewhattypeofbloodshouldbegivenAntibodyscreenand/orIDTheantibodyscreenwilldetectthepresenceofanyunexpectedantibodiesinpatientserumIfantibodiesaredetected,identificationshouldbeperformedusingpanelcells(withanautocontrol)IS37°(LISS)AHGIfanantibodyispresent,unitsnegativefortheantigenmustbegivenProceedtothecrossmatch…CrossmatchingPurpose:PreventtransfusionreactionsIncreaseinvivosurvivalofredcellsDoublechecksforABOerrorsAnothermethodofdetectingantibodiesCrossmatchTwotypesofcrossmatchesMajor–routinelyperformedinlabsMinor–notrequiredbyAABBsince1976HistoryMajorvsMinorCrossmatchWhyistheminorcrossmatchunnecessary?DonatedunitsaretestedforantibodiesMostbloodistransfusedaspackedcells,havinglittleantibodiesCrossmatchesTheAABBandFDAdevelopthestandardsforbloodbankingAccordingtotheAABBStandards:

Thecrossmatch“shallusemethodsthatdemonstrateABOincompatibilityandclinicallysignificantantibodiestoredcellantigensandshallincludeanantiglobulinphase”Involveslabeling,inspecting,andissuingthebloodunitSpecialprocedures/protocolsmustbeavailablesothattheautologousunitisfoundandtransfusedtotherecipientHemolytictransfusionreaction,ifthepatient'santibodyistooweaktobedetected.Weakornonreactedwithanyorsomecellreagent.PhenotypingforsomeRBCsantigensAgsdeteriorateonstorage(Duffy,P1,M,Lewis)Aneonatewhois<4monthsolddoesnothaveantibodiesUnlikelytocausesignificantdestructiontorecipient'sRBCs.RTandISarenotrequired.AntibodydirectedagainstantigenonscreeningcellAccordingtotheAABBStandards:DetectallantibodiesMinor–notrequiredbyAABBsince1976Compatibilitytesting?DetectABO/RherrorsDonorRBCs(washed)CrossmatchDonorRBCs(washed)PatientserumNoagglutination~compatibleAgglutination~incompatibleTheprocedureDonorcellsaretakenfromsegmentsthatareattachedtotheunititselfSegmentsareasamplingofthebloodandeliminatehavingtoopentheactualunitAgglutination~incompatibleMostABOincompatibilitiesUnlikelytocausesignificantdestructiontorecipient'sRBCs.CrossmatchProcedureEmergencyReleaseExpiredreagentshouldnotbeused.PlasmacompatibilitytableAntibodydetection/identificationInfusionDeviceInvolveslabeling,inspecting,andissuingthebloodunitWhatcanbegiveninanemergency?AHGabsence,inadequacy,orneutralizationProblemsEncounteredInaddition,thelabmaycrossmatchadditionalunitsasaprecautionifmorebloodisneededCrossmatches…UnitsofwholebloodwithsegmentsattachedCrossmatchProcedureifantibodiesareNOTdetected:Onlyimmediatespin(IS)isperformedusingpatientserumanddonorbloodsuspensionThisfulfillstheAABBstandardforABOincompatibilityThisisanINCOMPLETECROSSMATCHIfantibodiesAREdetected:AntigennegativeunitsfoundandX-matchedAllphasesaretested:IS,37°,AHGThisisaCOMPLETECROSSMATCHCrossmatches…WillVerifydonorcellABOcompatibilityDetectmostantibodiesagainstdonorcellsWillNotGuaranteenormalsurvivalofRBCsPreventpatientfromdevelopinganantibodyDetectallantibodiesPreventdelayedtransfusionreactionsDetectABO/RherrorsIncompatiblecrossmatchesAntibodyscreenCrossmatchCauseResolutionPosNegAntibodydirectedagainstantigenonscreeningcellIDantibody,selectantigennegativebloodNegPosAntibodydirectedagainstantigenondonorcellwhichmaynotbeonscreeningcellORdonorunitmayhaveIgGpreviouslyattachedIDantibody,selectantigennegativebloodORperformDATondonorunitPosPosAntibodiesdirectedagainstbothscreeninganddonorcellsAntibodyID,selectantigennegativebloodAdditionalInformationonTypesofCompatibilityTestsManual(ISandIAT)GelTechnologyElectronic(Computerized)CrossmatchRedcellAffinityColumnTechnology(ReACT)SolidPhaseAdherenceAssays(SPAA)GelTechnologyPatientserum,and1%ofsuspendedRBCsinLIMaredispensedintothemicrotubeandincubatedat37oCfor15minutes.Thecardcontainingthemicrotubesisthencentrifugedatacontrolledspeedfor10minutes.Atthestartofcentrifugationthecellsareseparatedfromtheserum;thentheymeettheAHGcontainedinthemicrotube.Finallythecellsaretrappedbythegel(ifagglutinated)orpellettothebottomofthetube.XMatchPhasesPHASEPURPOSEDETECTSISdetectionIgMcoldagglutininMostABOincompatibilities37oC,LISSIgGAbsPotentRhAbsAHGIgGAbsRh,Duffy,Kidd,othersOCCAHGabsence,inadequacy,orneutralizationSensitizedRBCs`clinicallyinsignificantsuchasanti-M,-N,-Lea,-Leb,and-I.

Thisphaseisusuallyreadonlymacroscopicallyforagglutination

ThisphaseisreadmicroscopicallyforagglutinationaddedtoallnegativetestsandmustproduceapositiveresultLimitationsofPretransfusionTesting

Hemolytictransfusionreaction,ifthepatient'santibodyistooweaktobedetected.Standardantibodydetectionmethodssuchastheindirectantiglobulintestrequireseveral100antibodymoleculesperredcelltoproducedetectablereactions.Ahemolytictransfusionreactionduetopatientmisidentification.Forexample,groupAredcells(meantfortransfusiontoagroupOrecipient)willbecompatibleinvitrotestswithanincorrectspecimendrawnfromagroupAperson.LimitationsofPretransfusionTestingHemolytictransfusionreactionifdonorredcellsareinadvertentlyhemolysedbeforeenteringthepatient,e.g.,redcellshemolysedbyanimproperlyfunctioningbloodwarmerorredcellshemolysedbycontactwithanicepackinatransportcontainer.Nonhemolytictransfusionreactionssuchasallergic,febrile,andotherreactions.Pretransfusiontestaremeanttodetectonlyredcellantibodies.IncompatiblecrossmatchABOincompatibility

RecheckpatientandbloodunitABOgroupClinicallySignificantAbDAT&IATAntibodyDetectionIATIATisusedtodetectclinicallysignificantIgGantibodiesboundtoRBCSinvitro.DetectionoffreeAbsinpatient'sserum.ClinicallysignificantIgGAbscausehemolysisorreducesurvivaloftransfusedRBCs,likeD,c,E,K,k,JKa,JKb,FYa,FYb,S,s.UnimportantAbs:I,P1,Lea,Leb,M,N.ThoseAbsareUnimportantduetoMostlycoldreactive(IgM)CouldbeneutralizedIAT

isprincipallyusedforAbdetectionAbinvestigation(identification)AbtitrationCompatibilitytestingPhenotypingforsomeRBCsantigensReagentusedinIATCharacteristicsofRBCsreagent:Threecellreagentsets,SerocyteI,II,III.RBCsantigenscorrespondingtoimportantandclinicallysignificantAbs.Thereagentcellsaresuspendedinsalineand2%-5%antibiotic.Expiredreagentshouldnotbeused.Anantigramdefiningphenotypeofreagentcellmustincludedwitheverycell.Allreagentshouldbestoredat1-6oC.

PerformanceIATMostclinicallysignificantAbsreactat37oC,RTandISarenotrequired.AHGisusuallymonospecificIgGNoneedforpolyspecificAHG.BecauseAntiCwilldetectAbsduringtheincubationphase.AntiCenhancethedetectionofcoldagglutininsuchasantiI,antiPthatarenonsignificantAbsandnotimportantintransfusion.Delayingtransfusionincriticalsituations.InterpretationResultofIATNegativeIATOCCPositive:ReportIATnegativePositiveIATPerform:Abidentification.Abtitration.ItsnotalifethreateningsituationwhenanunexpectedclinicallyimportantAbstoanRBCsAgsinabloodunitisnotdetected,BecauseTheplasmavolumeissmall,andAbswillbedilutedinrecipientcirculationUnlikelytocausesignificantdestructiontorecipient'sRBCs.AntibodyIdentification0.3%-2.8%ofthepopulationareAbmakers,theyproduceclinicallysignificantAb.PregnancyandtransfusionarethecommoncauseofimmunizationtoRBCsAgs.SerologicalPropertiesAbsofbloodgroupsvaryinimportanceandsignificanceduetoserologicalpropertiesTemperaturephase37oCreactiveAbsasantiD,c,E,K,k,JKa,JKb,FYa,FYb,S,s.ColdagglutininasantiLea,Leb,I,P1,M,N.InducingHDNandHTRActivationcomplement.NeutralizingbysolublebloodgroupAgsProblemsEncounteredAutoantibodydirectedagainstowncellAgs

solutionElution

IgGAbscanbedissociatedfromRBCsmembraneAgsbyphysicalorchemicalmeans,someproceduresdestroymembrane,someleaveitintact.Supernatantfluid(elute)containtheautoantibodieswhichcanbeidentified..

ProblemsEncounteredCombinationofautoandalloantibody

solutionTreatmentwithZZAP(Adsorption)ZZAPeffectivelyremovesautoantibodies(digestion)allowingmorecompleteabsorptionofautoantibodiesfrompatient'sserum,allowingdetectionandidentificationofalloantibody.ChemicallytreatedallogenicRBCsTreatedRBCSwithproteolyticenzymes,altersomeAgsProblemsEncounteredWeakornonreactedwithanyorsomecellreagent.VariablereactionVariableexpressionofAgs(P1,Lewis)Agsdeteriorateonstorage(Duffy,P1,M,Lewis)AbsisreactingwithmorethanoneAg.DDCCee

3+Ddccee1+

SolutionUsedoubledoseofcorrespondingAgs,(Rh,Kidd,Duffy,MNS)UsenewcellreagentPost-analyticalphasePost-analyticalphaseInvolveslabeling,inspecting,andissuingthebloodunitLabelingformincludespatient’sfullname,IDnumber,ABO/Rhofpatientandunit,donor#,compatibilityresults,andtechIDFormisattachedtothedonorunitandonlyreleasedfortherecipientTheunitisvisuallyinspectedforabnormalities,suchasbacterialcontamination,clots,etcBacterialcontaminationThisunitshowsbacterialcontaminationandshouldNOTbegiventothepatientTheplasmainthesegmentsisfine,buttheplasmaintheunitshowsheavyhemolysisfrombacteriaIssuingbloodWhenit’stimetoreleaseabloodproducttothenurseorphysician,afew“checks”mustbedoneRequisitionformComparingrequisitionformdonorunittagbloodproductlabelNameofpersonsissuingandpickingupbloodDateandtimeofreleaseExpirationdateWhatiftheunitisunused?BloodcanbereturnedifitisnotneededfortransfusionUnitclosurehastoremainunopenedStoragetemperaturemusthaveremainedintherequiredrange(1°to10°CforRBCs)Ifnotatcorrecttemp,unitmustbereturnedwithin30minutesofissueInfusionDeviceBloodWarmerSpecialCircumstancesEmergencyReleaseInanemergency(ERorOR),theremaynotbeenoughtimetotesttherecipient’ssampleInthiscase,bloodisreleasedonlywhensignedbythephysician(Onegative)ThetagmustindicateitisnotcrossmatchedSegmentsshouldberetainedforX-matchingEverydetailisdocumented(names,dates..)EmergencyReleaseOncethespecimenisreceived,ABO/RhtypingandantibodyscreeningshouldbeperformedCrossmatchingthesegmentsfromthereleasedunitshouldbetestedInaddition,thelabmaycrossmatchadditionalunitsasaprecautionifmorebloodisneededIfdeathshouldoccur,testingshouldbecompleteenoughtoshowthatthedeathwasunrelatedtoanincompatibilityWhatcanbegiveninanemergency?GroupORh-negativeredcellsorABplasmaEmergencyreleaseWomenbeloworofchildbearingageOrifindoubtGroupORh-positiveredcellsUsedasasubstitutionMaleorelderlyfemalesAppropriateunitstogiveABOcompatibleshouldalwaysbegivenfirstPatient’sType1stChoiceOtherChoicesOONoneAAOBBOABABA,B,OGroupOindividualsare“universaldonors”,theycandonatetoanybloodgroupbecausetheyhavenoAorBantigensGroupABindividualsare“universalrecipients”,theycanreceivebloodfromanygroupbecausetheydonothaveAorBantibodiesRedbloodcellcompatibilitytable

AB+

AB-

B+

B-

A+

A-

O+O-AB+AB-B+B-A+A-O+O-DonorRecipient

Plasma

compatibilitytable

O

B

A

ABABBAODonorRecipientNeonatalTransfusionAne