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'\  `Roman PSX'  8. DAT/Elution Study 2lab8 (Carolyn Ragland 0 .  . Rh Phenotyping ; Task To type three (3) cell samples for the following Rh antigens: D, C, E, c ! , o ; Aim To determine the Rh phenotype and probable genotype of three (3) cell samples. ; Introduction In this procedure, commonly tested antigens of the Rh system will be studied. Normally the only Rh antigen identified in routine pretransfusion testing is the D antigen.XU6Non-ProfitNon-Profit Stamp#$RB>z  0 @X<<<<@ExLLx d'dxd Level 1 Level 2 Level 3 Level 4 Level 5(?2fF$ SY!       ($     (?2fF$ SY!       " !C<< c A "fF"       @XX%w @@ NonProfitOrg.  @PP U.S.Postage@ PAID@uu Permit2757  @EE Austin,Texas.D4`P,BoardBoard of TrusteesC'.D4A`ArialTTC BoardofTrustees -%.D4`-Dr.BruceM.Murray,ChairmanDellaMayMoore,ViceChairmanRooseveltLeaks,SecretaryJanAlbersPeteFosterLindaGrayMackRayHernandezLawrenceE.JenkinsMurrayShawDr.DanAngel,President.D4A`ArialTTT}0HeadingFancy ACC Heading    xxxW)64 *Times New RomanTTW@.. AustinCommunityCollege  6*'Fd66*'Fd66*'t"Fd664 *Times New RomanTTP:FTFA MinutesFirst Pages%Xf$ hN p x (#%'0*,.8135@8:<H?AXX8(1Xdd8  @  FULLTIMEFACULTYASSOCIATION  @+Minutes   , _ -fF'       (1XXXs%Xf$ hN p x (#%'0*,.8135@8:<H?AX J(# Minutes   ?*' xdEx?(?2fF$ SY!       ^>8ACC AddressACC Return AddressAustinCommunityCollegeP.O.Box140526Austin,Texas78714dC,EqualEqual Opportunity Statement@.. ACCisanequalopportunityinstitution. N24 labels4 line labels ,  , ,       \RA'*ty CollegeInternal Audit Department\ X X R& 8BibliogrphyBibliography0....fE2Doc InitInitialize Document Style3( 7   (456($0 ($0 0 (@$0  0` (#(#0 ` (#` (# `   A_ekqwDocumentDocument StyleI.1.A.a.(1)(a)i)a)jD4Tech InitInitialize Technical Style( 9 ( CuyTechnicalTechnical Document Style11.11.1.11.1.1.11.1.1.1.11.1.1.1.1.11.1.1.1.1.1.11.1.1.1.1.1.1.1x?;u2PleadingHeader for numbered pleading paper ;  <fF(       (1XXXXX?*'*dE*??*'*dE*?\\1\\2\\3\\4\\5\\6\\7\\8\\910111213141516171819202122232425262728  .+(?2fF$ SY!         -fF'       _HH>XGXXX >X>XGJ1>EXERCISE8&9#>J1#J1>#JXZXJ1#@@:$LaboratoryProcedureManual  X #>XGXXJXZ#A*)  xd2EHCH!A'6X'A`"Sans Serif PS ) @fF)       _HH>XGXXX?*' XxdEHWH!? 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XX,8YXXdd8LLExercise8 0 P DirectAntiglobulinTest(DAT)P $P $ Exercise90 P ElutionStudy `P $P $ LZL (H$H$(Task  + ToperformtheDATandelutionprocedurewithcorrectinterpretationofresults. + (H` H` ( C  (H$H$(Aim  /  Toperformwith100%accuracytheDATprocedureandelutionprocedurewithcorrect /  interpretationofresults.   (H` H` (    (H$H$(Introduction   t  DAT   t Thedirectantiglobulintest(DAT)isusedtodetect invivoredbloodcellsensitization .  L  Washedredbloodcellsfromthepatientaredirectlytestedwithantiglobulinserum.The 8  specimenofchoicefortheDATisan_EDTA_Ԁspecimen.Ifaclottedspecimenisuseda $  falsepositiveresultmaybeobtainedduetononspecificbindingofcomplementtothe   redbloodcellsinvitro._EDTA_Ԁbindscalciumwhichisnecessaryforcomplement {  activation.Thefocusofthislabistodeterminesensitizationofbabysredbloodcells, g  buttheDATisaprocedurewhichmayalsobeperformedonothertypesofpatientsas S wellasdonorunits. ?  TheDATisutilizedinthebloodbanktoaidinthediagnosisofavarietyof  conditionssuchas:   1.0 Diagnosisofhemolyticdiseaseofthenewborn (_HDN_).Apregnantwoman T mayform_IgG_Ԁantibodiesdirectedagainstantigensonbaby'scellswhileitisin @ _utero_.Thesemayhavetheabilitytocrosstheplacentaandattachtothebaby's , redbloodcells._IgG_Ԁistheonlyantibodyclasscapableofcrossingtheplacenta. T$ T$ 2.0  Diagnosisofautoimmunehemolyticanemia .Apatientformsantibodies o directedagainsttheirownredbloodcellscausingdecreasedcellsurvival.[ T$ T$ 3.0  Investigationofredbloodcellsensitizationcausedbydrugs .Drugs 3  sometimesinducetheformationofantibodies,eitheragainstthedrugitselfor ! againstintrinsicredbloodcellantigens.Drugsactas_haptens_Ԁelicitingantibodies  " onlyaftertheyhavebeenfirmlyboundtoaproteincarrier.Onceformed,the "x antibodiescanreactwiththesmall_hapten_Ԁindependentofanyprotein #d  attachment.Drug-inducedantibodiesmaycausepositive_DATs_Ԁandsometimes $P!! hemolytic_anemias_.%<"" T$ T$ &  4.0  Investigationoftransfusionreactions .Apatientmayforman_tibodies_Ԁagainst '$$ transfusedcells,causingthetransfusedcellstobecoatedwithantibodies (%% in-vivo.k)%& T$ T$ ''fIfapatienthasapositiveDAT,anelutionprocedureshouldbeperformed.Acritical C+'( partoftheelutionprocedureincludesthepatienthistory.Thepatient'sdiagnosis, /,() transfusionandpregnancyhistory,andrecent/currentmedicationsare vitalpieces -)* ofinformation inworkingupapositiveDAT.  .*+  .|+,  Elution   Thepurposeofanelutionistocausethereleaseofantibodymoleculesfromthered W bloodcellmembrane.Oncefreeinsolutionthe_eluted_Ԁantibodiesaretestedagainst C reagentredbloodcellstodetermineifanimmuneantibodyspecificityispresent. / Antibodyspecificityisexpectedincasesof_HDN_,transfusionreactionsduetoimmune  antibodiesandsometypesofautoimmunehemolytic_anemias_._Eluates_Ԁareusuallynon   reactiveincasesofdruginduced_DATs_.  t Antibodyrelease(elution)canbeaccomplishedeitherbydisruptingtheantibodyorby  L  changingconditionstofavordissociationofantibodiesfromantigen.Manytechniques  8  areavailable.Nosinglemethodisbestinallsituations.Ifaneluatepreparedbyone  $  techniquegivesunsatisfactoryresults,itmaybehelpfultoprepareanothereluatebya   differenttechnique.Negativeresultsinanantibodyscreeningtestdoneonaneluate {  doesnotmeananantibodyisabsent;theantibodymayreactwithalowincidence g  antigenorwithadrug. S  Thenexttwo(2)laboratoriesarebloodbanktestsusedintheevaluationofhemolytic + diseaseofthenewborn(_HDN_).Immuneantibodiesarestimulatedasaresultof  exposuretoredbloodcellantigenswhichanindividualdoesnotpossess.Ifawoman  hasbeenexposedtoforeignredcellantigens(mostcommonlyasaresultoftransfusion p orpreviouspregnancy),shemayformimmune(_IgG_)antibodiescapableofcrossingthe \ placentaandattachingtoantigenpositivefetalcells.Thiscausesthebaby'sredblood H cellstobecoatedwithantibodyanddestroyed.Theresultingeffectsonthechildare 4 collectivelytermedhemolyticdiseaseofthenewborn(_HDN_).Theseverityofthe_HDN_   variesandisdependentupontheantibodyinvolved.Themostsevere_HDN_isduetoD   negativemotherswhohaveformedimmuneanti-Dthatcrossestheplacentaand w destroystheDpositivecellsofherDpositivefetus.Ifthe_HDN_Ԁissevere,thebabywill c needan exchangetransfusion withbloodcompatiblewiththebabyandthemother.) O Themildestandmostcommonformof_HDN_ԀisduetomaternalABOantibodies.This ; isdetectedingroupAorBinfantsofgroupOmothers._Phototherapy_Ԁisthetreatment ' ofchoiceforABO_HDN_.Antibodiesto otherbloodgroupantigensmaycause   varyingdegreesof_HDN_.     !l!  Principle  "X!  Part1 ofthislabistodeterminewhetheraninfant'sredbloodcellshavebeensensitized "X! bymaternalantibodybyperformingthedirectantiglobulintest(DAT).Please #D " rememberthattheDATisageneralprocedurethatisperformedonbabies,adults $0!# anddonorunits.Thepurposeofthetestinthislaboratorysessionistodetect %"$ sensitizationofbabycellsbymaternalantibody,butthistestisalsoapplicableinmany &#% othersituations. s'#& TheDATproceduredetectsin-vivosensitizationofredbloodcells.Ifanegativeresult K)%( isobtained,nosensitizationhasoccurred.Apositiveresultindicatesin-vivo 7*&) sensitizationhasoccurredandfurthertestingisrequired. #+'* TheDATprocedureisroutinelyperformedonthecordbloodsofnewborns. ,|),  .T+. _Thespecimenfortestingisobtainedfromthebaby'sumbilicalcordimmediately  followingbirth.Asyringeisusedtowithdrawaspecimentopreventcontamination k withWharton'sjelly.Wharton'sjellyisagelatinoussubstancewhichcoatstheumbilical W cord.Ifthecordspecimenbecomescontaminatedwiththissubstance,itwillcause C non-specificagglutinationofthecells(falsepositives).Falsepositivereactionsare / usuallydiscoveredwhentheABO/Dtypingisperformed.Allforwardtypingtubesare  positive,anRhcontrolisrun,anditisalsopositive,invalidatingthetest.Itisawater   solublesubstance,andonceitisdiscoveredasacontaminant,canberemovedby8-10  t washeswithlargevolumesofsaline.Thecordspecimenwillbecontaminatedifthe  ` specimeniscollectedbycuttingthecordandallowingthebloodtodripinthetubeor  L  ifthebloodis milkedintothetube.  8  WhenapositivecordbloodDATisobtainedaseriesofstepsarerequiredtoidentifythe   offendingantibody.Thefirststepindeterminingtheidentityofthecoatingantibody {  istoperformaTypeandScreenonthemomandaABO/Dtypingontheinfant.Please g  rememberthatareversetypecannotbeperformedonbabies,astheABOantibodies S  presentintheirserum/plasmaisofmaternalorigin.Theresultsoftheantibodyscreen ? onthemotherwilldeterminewhichtypeofelutiontoperform,assomeelution + proceduresarebestatrecoveringalloantibodies,whileothersarebestatrecovering  immuneABOantibodies.Ifthemother'santibodyscreenispositive,apanelstudyis  doneonthemother'sserum.Anacidelutionmustbeperformedontheinfant'scord p cellstoverifythattheantibodyinthemother'sserumis,indeed,coatingthebaby'scells. \ Ifthemom'sscreensarenegativeandsheisABOincompatiblewithherinfantthena H heatelutionisperformedontheinfant'scordcellstocheckforanABOantibody. 4  Part2 ofthislabwillinvolvetheperformanceofanelutionontheinfant'scordcellsto   determinethespecificityofthematernalantibodycoatingtheinfant'sredbloodcells. w Inanelutionprocedure,thecellsarewashedfreeofallunboundantibodyanda c procedureisperformedonthecellstocausethereleaseoftheantibodymoleculesfrom O thecellsintoasolution.Theeluateisthentestedagainstreagentredcellstodetermine ; thespecificity.Rememberthatanyantibodyrecoveredintheeluatepreparedfromthe ' infant'scellsisof maternalorigin .   TodayDATswillbeperformedoncordbloods.Ifapositiveresultisobtained,perform !p! aTypeandScreenonthemomandanABO/Dtypingonthecordcells.Fromthis "\" informationdeterminethetypeofelutiontobeperformed.Nextweektheelution #H # procedurewillbedoneandtheeluatetestedtodeterminethespecificityoftheantibody $4!$ coatingthecells. % "%   -l*.  PleasekeepthefollowingflowcharthandyasyouperformthecordbloodDAT   testing.  g  O   &Y"%XXA& %"%&Y=p<d<R!H` H` = H` H` H H  H H H` H` wDC3/Z ,8@ H@E88w  @ DATresults ; @ oncordcells  {DE51Z ,8@@E88{   ; 4p<d<RAH H 4 H H H$H$ l  $$H H  H$H$$$4p<d<RAH$H$4 H H H$H$@ NEGATIVE      @''.POSITIVE      yDE51Z ,8@@Es 8s 8y@ Noelution  |  @ necessary  T   s   {DE51Z ,8@@Es 8s 8{  @  *PerformTypeand  |  @~~,Screenonmom  T  {DE51Z ,8@@E88{    @@'PerformABO/Dtypingon 3  cordblood      99@cc*Rhoncordcells   s  4p<d<RaH H 4 H H H$H$L8k4AH H 4 H2 H2 H H  8 wDC3/Z ,h@ H@Edhdhw@0 0 Ifmom'sscreens g @ arepositive ? yDE51Z ,h@@Edhdhy@ Performacidelution H @# # oncordcellsand   @  testeluateagainst w @  panelcells O  yDE51Z ,h@@Ed'hd'hy@ Testmom'sserum X# @ againstpanelcells 0$   %  8% {DE51Z ,h@@E,h,h{   , ,  , ,   @77:Ifmom'sscreens g' @CC:arenegativeand ?( @<infantisABO ) @:incompatiblewith p* @Amom H+    , , {DE51Z ,h@@E,h,h { , ,   @8Performheatelution O. @9oncordcells;test '/ @8eluateandlastwash 0 @9againstAcells,B X1 @UU:cellsandscreen 02 @@cells  3  _!4 4@H2 H2 4 7"4 AXZX %A&XYXXAXZLH4X<0f>tXH ( @H$H$(DAT .`+C  Procedure     3 A X X<6631.0 Labelatubeforeachcordspecimenusingthemother'sfullnameandhospital .`+ number. T$ T$  NOTE :Assoonasyouhavelabeledthetubeplaceimmediatelypickuptheappropriate 2 cordbloodandplacetherbcsinthelabeledtube.Placingthecellsinthetubeafter j verifyingitisthecorrectsampleimmediatelyafterlabelingwillhelppreventyoufrom  puttingthewrongcellsinthewrongtube. M 2.0 Prepareawashed2-5%suspensionofcordcellsinaproperlylabeledtesttube.  T$ T$ 3.0 Placeone(1)dropofcellsuspensiontobetestedinaproperlylabeledtesttube. 2 T$ T$ 4.0 Washthree(3)timeswithsaline,decantingcompletelyaftereachwash. d T$ T$ 5.0 Afterlastwash,blottubewithgauzetoobtainadrycellbutton.  T$ T$ 6.0 Addtwo(2)dropsofAHGreagent.G  T$ T$ 7.0 Gentlyshaketubetomixandcentrifugefor10-15seconds.y  T$ T$ 8.0 Examineforagglutinationmacroscopicallyandmicroscopically.,  T$ T$  9.0 AddonedropofIgG!sensitizedredbloodcells(checkcells)toanytubes ^  whichhavebeenrecordedasnegative.  T$ T$ 10.0 Gentlymix,centrifugefor15seconds.A T$ T$ 11.0 Resuspendthecellbutton,gradeandrecordresultsandinterpretation.s T$ T$ 0 NOTE: Iftheinitialwashingadequatelyremovedallunboundglobulin,the & antiglobulinserumwillstillbereactiveandcapableofagglutinatingthe ^ sensitizedcontrolredbloodcells.Thus,thenegativetestresultobtainedforthe  patientcanbeconsideredvalid.Thisisanimportantcontrolbecauseitisthe A onlywaytomonitortheadequacywithwhicheachtestiswashed. s T$ T$ ( @H4H4( &  (,2@H$H$(Interpretation  X 1.0 Negative,noin-vivosensitizationdetected.X T$ T$ 2.0 Positive,in-vivosensitizationdetected;elutionstudiesshouldbeperformed.   Thetypeofelutiondonewillbedeterminedbytheresultsofthemother's ;  typeandscreen.*Atthistime,performtheTypeandScreenonthe m!  maternal specimenaccordingtoyourpreviousexercise."  T$ T$ 0 Whilethetypeandscreenonthemotherisincubating,performanABO/D %! onthe baby's cells.Areversetypecannotbeperformedonaninfantsince 5&" allantibodiesinthebabyareofmaternalorigin.Afterallresultsarein,look g'# attheflowcharttodeterminewhicheluatewillbeperformednextweek. (% T$ T$   HeatElution 7+' Introduction  ,$)    Theredbloodcellsforanyelutiontechniquemustbethoroughlywashedtoremove *~'! allantibodyexceptthatboundtothecells.Sixtoeightwasheswithlargevolumes /,(" ofsalineareusuallysufficient.*Adequacyofwashingisdeterminedbytestingsaline a-)# fromthelastwashagainstreagentscreencellsforthepresenceofantibody.If .+$ positivereactionsareobtainedwiththelastwashthisisanindicationthatthered  bloodcellswerenotwashedsufficientlyandthattherearestillcontaminating 2 serumantibodiespresent.Thiswouldinvalidatetheresultsoftheeluateandthe d procedurewouldneedtoberepeatedusingamorethoroughlywashedrbcsample.  Theheatelutiontechniqueisusedtoremoveantibodiesfromredbloodcellscoated y  in-vivo.ItisusedprimarilytorecoverandidentifyABOantibodiesincasesof  , hemolyticdiseaseofthenewborn(HDN)andinpatientswhohavebeentransfused  ^ withnon-ABO-identicalbloodorbloodcomponents.Thecoatedredbloodcellsare   thoroughlywashed,analbuminsolutionisaddedandthecellsareheatedto56C. A  Thisamountofheatwillcausedisruptionoftheantigenantibodybond,theantibody s  will popoffthecoatedrbcsintothealbuminsolution.Aftersufficientheatingthe &  solutioniscentrifugedandthesupernatantfluid(eluate)isremoved.Thesupernatant X  willbeveryhemolyzedandyoumayneedalamptoprovidealightedbackground    toviewtheeluate/rbcinterface.Contaminatingtheeluatewithrbcsmaycausefalse ; positivereactionsinthetest.Thesupernatantfluidwillcontaintheantibodiesthat m werecoatingthecells.Theeluateistestedagainstreagentredbloodcellsto   determinethespecificity. R  1V,2`H H 1Reagents  +( 1.0 Seepage1*r'- T$ T$ 2.0 6%bovinealbumin,preparedbydiluting22%or30%bovinealbuminwithsaline#,(. T$ T$ 3.0 CordbloodredbloodcellsfrompreviouslabU-)/ T$ T$ 4.0 56$Cwaterbath .+0 T$ T$ (V@HH(   (w@H$H$(Preparing 2 Eluate   1.0 CentrifugethesamplewiththepositiveDAT.Transferthesupernatantserumor 2 plasmatoaseparate,properlylabeledtube.d T$ T$ 2.0 Place20dropsoftheredcellsinaproperlylabeled12x75mmtube. T$ T$ 3.0 Washthecellseight(8)timeswithlargevolumesofsaline.G T$ T$  NOTE : DONOT decantthesalineoffthecells,thevolumeofcellsusediscritical. y  ЀRemovethesupernatantsalinewithapipet.  2 4.0 Mixcells vigorously betweenwashesbyaddingasmallvolumeofsalineand  d thumpingthetubevigorouslywithfingerorcoveringthetubewithparafilmand   invertingthetubeuntilallcellsareresuspendedoffthebottomofthetube.Then, G  completefillingoftube,addingsalinevigorously(donotoverfill,mayneedto y  parafilmtopoftube).,  T$ T$  NOTE :Ifcellsarenotcompletelyresuspendedbetweenwashesantibodymaybetrapped ^  inbetweenthepackedrbcs,thiswillcauseafalsepositivereactioninthelastwashaswell  astheeluateandtheprocedurewillneedtoberepeated. G 5.0 Putthesalinefromwash#8inatubelabeledwithpatientnameand LAST y WASH .2 T$ T$ 6.0 Add20dropsof6%bovinealbumintothewashedpackedredbloodcells.j T$ T$ 7.0 Placethetubesat56$Cfor10minutes,agitatingthetube constantly duringthis  timewithtwoapplicatorsticks.M T$ T$ 8.0 Centrifugethetubeat1,000Xgforfive(5)minutes. T$ T$ 9.0 Immediatelytransferthesupernatanteluateintoacleantesttube,andtestin 2 parallelwiththefinalwashsupernatant.d T$ T$  ", (w@HH( #^   (w?@H$H$(Testingthe  +'! Eluate  u,(" 1.0 Label10,12x75testtubes(eluate( EL )=A1,B,S1,S2,andS3;lastwash( LW )  +'" =A1,B,S1,S2,andS3)andpatientinitials.A,(# $ $ 2.0 Addone(1)dropofthereagentredcellstoeachoftheproperlylabeledtubes.s-)$ $ $ 3.0 Tothefive(5)tubeslabeled EL,addthree(3)tofour(4)dropsoftheeluate. .&+% $ $ 4.0 Tothefive(5)tubeslabeled LW,addthesamenumberofdropsofthelastwash  aseluate,IE,ifonly3dropsofeluatewereadded,placeonly3dropsoflastwash 2 intheLWtubes.d $ $ 5.0 Mixandincubateat37$for30minutes. $ $ 6.0 Removefromheatblockand immediately washthree(3)timeswithsaline, G decantingthoroughlybetweenwashesandblottingthetubesafterthelastwash.y  $ $ 7.0 Addtwo(2)dropsofantiglobulinreagenttoeachtube,mixgentlyandcentrifuge  , for15seconds. ^ $ $ 8.0 Gentlydislodgecellbuttoncompletely.  $ $ 9.0 Readmacroscopicallyandnegativetubesmicro.A  $ $ 10.0 Recordresults;gradingreactions.s  $ $ 11.0 Addone(1)dropCoombscontrolcellstoallnegativetubes,centrifugefor15 &  secondsandreadforagglutination.Ifnoagglutinationispresent,testmustbe X  repeated.   $ $  ['# (w?@HH( (%  (c4@H$H$(Interpretation  *r'  1.0 If lastwashtubesarenegative,anyagglutinationofreagentcells *r'  witheluateindicatesrecoveryofantibodiesfromtheoriginalcells.#,(! ! ! 2.0 Ifthelastwashispositivewithanyofthetestcells,theoriginal U-)" cellsamplewasprobablyunderwashedandtheelution .+# procedureshouldberepeatedonamorethoroughlywashedcell  sample. 2 ! ! 3.0 Lackofagglutinationoftestcellsbytheeluateindicatesabsenceof d antibodiesspecificfortheantigensonthetestcells. ! ! 4.0 Inthecaseofcordblood,theinterpretationshouldbereportedas G follows: y  ! ! (c4@H H (  , 9% X<o!z]hA X9  P < ReactionswithCells 0 z P1] Interpretation  ^z$z$ A$XXYE0%$A  P"<A1P'BP)oS1Pxx-S2P0!S3   L  P#<+P'OP99*oOP.OP1!O z ImmuneantiAeluted  z    P#<OP'+P99*oOP.OP1!O z ImmuneantiBeluted Y    P"<+sP&+wP99*oOP.OP1!O z ImmuneantiAandantiA,Beluted :    P"<+wP&+sP99*oOP.OP1!O z ImmuneantiBandantiA,Beluted    P#<OP'OPoo*oP'(+onanycell)0 z Unexpectedantibody.Performpanelonmaternalserumyz$z$ - HXX<Y]h-EXZX%E00  0$$0P $$IfthemotherisgroupOandthebabyisgroupAorB,itisnotunusualtoobtain Z positivereactionswithboththeAandBcells.ThisreactivityisduetotheantiA,B F presentinthegroupOmothersserumwhichhascoatedthebabysrbcs.This 2 antibodywillreactwithbothAandBcells.P $P $ 0  0$$0P $$Usetheinterpretationchartabovealongwithyourcommonsensetointerpretyour u data.IfthemomisgroupOandthebabyisgroupA,itisimpossibletoelutean a immuneantiBfromthebabycellls.Thinkyourinterpretationthroughcarefully.MP $P $ 0  0$$0P $$ Fortheexamplebelow themomisgroupO,antibodyscreennegativeandthebaby % isgroupAwithapositiveDAT.Thefollowingelutionresultswereobtained.P $P $ *YZ[A ddd Xdd Xdd XH$H$,Sdd ,dd ,dd ,Xdd ,dd ,Xdd +  jj  6,!M "H 6A1cells 6,!M  "H 6Bcells 6,!M !"H 6S1 6,!M ""H 6S2 6,!M #"H 6S3 2(&M $"  H 2Eluate '!N% '4+ '!N&" '2+ ;1!N'" # ;0 ND%!N(" #  # N0 ND%!N)" #  # N0 1'%!N*" #  1LastWashControl ;1M#+ # ;0 ND%M#," #  # N0 ND%M#-" #  # N0 ND%M#." #  # N0 ND%M#/" #  # N05+)M#0" #   50  0$$0P $$Interpretation:ImmuneantiAandA,Belutedfromcordcells.U%!1P $P $   -'#3 XYXXEXZDiV FBzx? p @@E, ,, 0$Name_________________________________  0$Date_________________________________ W @( CordbloodDAT / @YY&RecordingResults      4  M,<&Y"%XXYA& %"%&Y'X HX'0$Name_________________________________ < XX0$Date_________________________________ / @ff' ElutionStudies  @&RecordingResults  # 0X pXc00  Mother'sName p  @p  $$ 0  HospitalNumber p  @p  \ $$ 0  Baby'sDAT  p O $$ 0  Baby'sABO/DTyping p  @p $$ 0  Mother'sABO/DTyping p l$$ 0  Mother'sAntibodyScreen p _$$ *NO[dA dSdd Sdd dd Xdd Xdd Xdd XYZH$H$,H@0 , @0 , @@ +  FH1 XdFCells UH@+?"Hd dUReactionswithEluate UH@+?"Hd dUReactionswithLastWashControl ]N6?" X  Hd X]A1 ;,!o," ; ?,!o," ? SD,o," X   XSB ;,!O  " ; ?,!O !" ? SD,O "" X   XSS1 ;,!/#" ; ?,!/$" ? SD,/%" X   XSS2 ;,!&" ; ?,!'" ? UD,(" X   XUS3 =,!)" = ?,!*" ?<20+" X    < AXZX %A&Interpretationofelutionresults:#A& %XAXZ#  p  #".   s'0&2 < XT8| pX P<0$Name_________________________________  - HXXT8DpD-$$U$ \& % 6A&Exercise80 P #A& % 6\&# 0 P $P $0 $ $ DirectAntiglobulinTest(DAT)$$ @&&'StudyQuestions  K AXZX %A&1.0  WhatdoestheDATproceduredetect?(1point)S $$ 2.0  Listfour(4)clinicalconditionsforwhichtheDATisuseful.(2points)\ $$ ]^@  ^@]3  .0    WhatisthespecimenofchoicefortheDAT?Why?(1point)^@͐]ی$$ Ќ  ^@  ^@]4  .0    WhatisthesourceofthespecimenusedfortheDATonnewborns?^@ʑ]ی_! $$ Ќ  ^@  ^@]5  .0    WhatisWharton'sjelly?Specifically,whatproblem(s)doesitcause?(2points)^@ʒ]ی%|"%$$ Ќ  ^@  ^@ē]6  .0    ListthethreetypesofHDNfrom mildest tomostsevereandstatetheantibodyspecificityinvolved G.*. ineach.(3points)^@ēߓ]ی3/+/$$ Ќ  ^@  ^@]7  .0    WhatantibodyclassisinvolvedinHDN?(0.5)^@:]ی `$$ Ќ  ^@  ^@ ]8  .0    StatetheadditionaltestingwhichmustbeperformedwhenaninfanthasapositiveDAT.Besureto {  statewhichtestisperformedonmomandwhichonbaby.(1.5points)^@ ']یg $$ Ќ  ^@  ^@]9  .0    WhatmustbedonefortheinfantiftheHDNissevereatbirth?(1point)^@] ی $$ Ќ  ^@  ^@]10  .0    Whycantareversetypebeperformedoninfants?(1point)^@] ی'$$ Ќ  .0   $$   7*&* 0$Name_________________________________  $$U$ `XZXAXZExercise90 P 0 P $P $0 $ $Elution#AXZX`XZE#ԀProcedureC$$ @'StudyQuestions  $ 1.0  Whatistheprincipleoftheelutionprocedure?(1point) \$$ 2.0  Listfour(4)piecesofinformationthatareusefulintheworkupofapositiveDAT.(2points)c $$ 3.0  WhatdetermineswhichtypeofelutionproceduretouseforanHDNworkup?(1point)0$$ 4.0  WhattypeofelutionisusedforABO-HDN?(0.5)7$$ 5.0  Intheelutionprocedure,whymustthecellsbethoroughlywashed?(1point)#@ "$$ &  6.0  Whatisthemostprobablecauseofpositiveresultswiththelastwashcontrol?(1point)3*&)$$ '3* .P+. Ї& t 7.0  Whatmustbedoneifthelastwashcontrolgivespositivereactions?(1point)$$ 'tb8.0  Whatdoeslackofagglutinationofthetestcellsbytheeluateindicate?(0.5) t$$ 9.0  Interpretthefollowingresultsobtainedinaheateluate.(1point){ $$ 0  BabyisApositiveS $$ 0  Momis0positive,screensarenegative ?$$ *acddH@0 H @0 @@ NOH$H$,Fdd ,dd ,dd ,dd ,Xdd ,Xdd +  $ $ '{ 'A1cells '{" 'Bcells '{" 'S1 '{" 'S2 '{" 'S3 {" Eluate  | 4+s '|" '2+ ;1|" # ;0 ND%|" #  # N0 ND%|" #  # N0 1'%|" #  1LastWash 2( { # 20 ND%{ " #  # N0 ND%{!" #  # N0 ND%{"" #  # N0 ND%{#" #  # N05+){$" #   50  Interpretation:%$$ 3 XT8| pHX310.0  WhenthemotherisgroupOandthebabyisAorBwhyisitcommontogetreactionsintheeluatewith 3) boththeAandBcells?(1point)*$$