《临床输血学实验》课件

TestingSpecimenRequirement2mlEDTAwholebloodFillinthetubelabel(s)withname

1精品医学临床输血学实验

TransfusionMedicine(bloodtransfusionlaboratories)上海市第六人民医院ShanghaiSixthPeopleHospital输血科TransfusionDepartment2精品医学HistoryAncienttimes敬畏血液-生命的源泉-饮血疗法Theperiod1500-1800治疗精神病血液有毒放血疗法-人体血液循环，为输血奠定基础-血液的携氧功能，合理治疗手段Thediscoveryofbloodgroups血型之父--Landsteiner3精品医学人类血型的发现，为安全输血提供重要保证。而且，在遗传学、人类学、法医学、免疫学、部分疾病的发病机制探讨上也具有重要意义。4精品医学Whilebloodcellantigens:RedcellantigensHLAantigensGranulocytes-specificantigensPlatelets（22）5精品医学

Partone

Redbloodcellgroup

目的要求：

掌握ABO血型的鉴定熟悉Rh血型的鉴定6精品医学

ABOSYSTEM

Principleofprocedure

红细胞上具有A抗原者为A型-RedbloodcellspossessAbloodgroupantigens有B抗原者为B型-RedbloodcellspossessBbloodgroupantigensA和B抗原都没有者为O型-RedbloodcellslackAandBbloodgroupantigensA和B抗原都有者为AB型-RedbloodcellspossessAandBbloodgroupantigens7精品医学RhSYSTEMThemostcomplexintheredcellsantigenIndividualswhopossesstheDantigenareRhpositive,wholackitareRhnegative.TheDantigeniscapableofstimulatingproductionofAnti-DinpersonslackingtheDantigen.Anti-DisaclinicallysignificantantibodycapableofcausingRBCdestructionandmayresultinhemolyticdiseaseofthenewbornandtransfusionreaction.8精品医学已发现的抗原40多个。涉及临床主要是5个抗原：C、c、D、E、e及其相应的特异性抗体。Fourotherantigens(C、c、E、e)accountforalmostalloftheRh-relatedtransfusionproblems.TheyarelessantigenicthanD,and/ortheantibodiesarelessclinicallydangerous.Buttheycancausethetransfusionreaction.9精品医学MaterialsRequiredCentrifuge0.9%salinepipetterreagents:Themonoclonalantibody(IgM)includeAnti-A

、Anti-B、Anti-D5%StandardRBC-Affirmagen10精品医学MethodsSolidphaseadherencetestTubetestGeltestManualSemi-automatedsystemAutomatedsystem11精品医学PROCEDURESpecimencollectionandpreparationRedbloodcellsuspensions(4%RBC)canbepreparedusingthefollowingcombinationsofsalineandpackedredbloodcells:SalineVolume---2mlPackedRBCVolume---100ul被检者RBC用生理盐水配成2-5%悬液待用(2ml0.9%NS+100ul压积红细胞）12精品医学TESTPROCEDURE---tubetest正定型：取小试管三只，分别用记号笔标记后加抗A、抗B、抗D血清各一滴，然后加受检者RBC悬液一滴。bloodgroupingAddonedropofAnti-A、Anti-B、Anti-DtothreetubesseparatelyAddonedropofa2-5%RBCsuspensiontothreetubesseparately13精品医学TESTPROCEDURE反定型：取小试管三只，分别用记号笔标记后加受检者血清一滴，然后加标准A、B及O型RBC各一滴。reversegroupingAddonedropofserum/plasmatothreetubesseparatelyAddonedropof5%standardRBCtothreetubesseparately14精品医学

将上述混匀后静置数分钟或3000r/m离心15s肉眼判断凝集度。Shakegentlytomixcontentsofreactiontubes.Centrifugesixtubeswith3000r/s,15m.Shakegentlywith45-60angles.Evaluatesixtubesforagglutinationand/orhemolysisupontestcompletion.Recordthereactionstrengthonthepaper.

15精品医学4+一个结实的大凝块，背景清楚。3+数个结实的凝块，背景清楚。2+中等大小的凝块加小凝块，背景清楚。1+小凝块，背景较浑浊。+或W+细胞有微小凝块，背景红色浑浊，通常用显微镜观察。MF混合外观凝集，少数红细胞形成较大的凝集块，而周围是游离的红细胞。0(negative)阴性，红细胞呈游离状态，无凝集象H(hemolysis)溶血，有游离血红蛋白。Evaluatetheagglutinationstrength16精品医学Anti-AAnti-BAnti-DAcBcOcgrouping+++———ARhDpositive—+++——BRhDpositive——+++—ORhDpositive+++———ABRhDpositiveInterpretationofresults17精品医学GroupinganomaliesColdauto-antibodiesAcquiredBUnexpectedmixedfieldreactionsPartialorweakDDnegativeIntrauterinetransfusions18精品医学RhDnegativeverification

Reagents:threedifferentAnti-Dhuman(IgG)monoclonalantibodyMethodIndirectantiglobulintest(IAT)Genetest

19精品医学IndirectantiglobulintestIncubatingthedonor’sredcellswiththeRhtypingseraWashingthecellsthreetimesAddingAHGCentrifugingthemixtureObservingitforagglutination20精品医学O有一种表现自己的性格,行动受感情左右,喜欢政治,好胜心强,对立斗争有戒心与喜欢交友并存,能看清形势,很注意别人对自己的好恶.有信念和理想.率直,具有现时与罗曼蒂克双重性.言行带有夸张性，讲话带有教育人和教训人的语气A内心想法不愿为人所知.对别人情绪及环境变化有敏锐反应,但对别人的想法缺乏客观理解.思想行动慎重而有规律,个性内向,没个性谨小慎微固步自封,遵从常理,重视家庭,富于计划性.擅长以说服和说明的调子讲话B开放型对人缺乏戒心,情绪善变,活动力强,不原受约束,行动奔放光明正大反复无常,富于灵活性,欢热闹场合.固执己见,善于批评,先下结论后作说明AB善于待人接物,很注意合情合理,期望在社会上施展能力,注意别人对自己的感情,厌恶表里不一.个性中庸,富有实践精神计划性强不喜欢指手划脚容易疲劳.表情不丰富,正式场合讲话呆板,回避感情激烈的讲话或事件TherelationshipbetweenABOgroupandpersonality21精品医学OABAB

ABOgroupandamativenessfemalemale22精品医学Parttwo

Redcellcompatibilitytesting-Crossmatch

目的要求：掌握交叉配血试验的原理熟悉交叉配血试验的操作23精品医学

CROSSMATCHING

Thecrossmatchisdefinedasaproceduretoexcludeincompatibilitybetweendonorandrecipient.Crossmatchingwillpickupincompatibilitiesbetweenthedonorandrecipientthatwillnotbeevidentonbloodtyping(asbloodtypingisnotavailableagainsteverybloodgroup,justthemajorones).Inaddition,thecrossmatchprocedurewillnotpickuplowtiterantibodiesandthuswillnotpreventdelayed-typehemolytictransfusionreactions24精品医学

Immediatespincrossmatch主侧配血---Majorcrossmatch(mostimportant)---comparingdonoreryghrocytestorecipientserumantibodiesinrecipientserumagainstdonorerythrocytes.次侧配血---Minorcrossmatch

Thiscomparesdonorserumtorecipienterythrocytesandchecksforpreformedantibodiesindonorserumthatcouldhemolyserecipientredcells25精品医学Choiceoftechniques盐水介质---saline酶介质---enzyme+liss抗球蛋白介质---IAT26精品医学

Crossmatchprocedure

Majorcrossmatch:Add1dof5%suspensionofdonorRBCsand1drecipientplasma/serumintothetube取受血者血清1-2滴+献血员2-5%红细胞盐水悬液1滴MinorcrossmatchAdd1dof5%suspensionofrecipientRBCsand1ddonorplasma/serumintothetube,取受血者2-5%红细胞盐水悬液1滴+献血员血清1-2滴27精品医学将上述混匀后静置数分钟或3000r/m离心15s肉眼判断凝集度。Centrifugesixtubeswith3000r/s,15m.Shakesoftlywith45-60angles.Evaluatetwotubesforagglutinationunderthemicroscope.28精品医学ResultWhenthereisanincompatiblereactiononthemajorcrossmatch,thedonorbloodshouldnotbetransfusionunderanycircumstances.Whenthereisanincompatiblereactionontheminorcrossmatch,thetransfusioncangoahead.However,ifthedonatedserumislikelytocontributesubstantiallytotheplasmavolumeoftherecipient,theserumshouldberemovedfromthedonorwholeblood.29精品医学SelectionofbloodRedcellcomponents---thesameABOandRhDgroupasthepatientIfABOidenticalbloodisnotavailable---groupORBC+groupABplasmaIfRhD-negativebloodarelimited---RhDpositivebloodmaybeselectedPatientwithclinicallysignificantredcellantibodyPatientwithautoimmunehaemolyticanaemiaMassivebloodtransfusion30精品医学

Partthree

Coombs’test

目的要求：掌握直接、间接抗人球蛋白试验的原理熟悉直接、间接抗人球蛋白试验的操了解直接、间接抗人球蛋白试验的临床应用31精品医学

一、抗人球蛋白试验的原理

倘若用提纯的或在人的全血清内的人的球蛋白给动物（如兔和山羊）注射，动物对外来的蛋白质将会产生抗体，即抗人球蛋白。32精品医学抗人球蛋白血清，在适当处理之后将会与人球蛋白发生特异性反应。如果球蛋白分子，或者抗体或补体附着于红细胞膜上，抗球蛋白血清将会与球蛋白结合。因为球蛋白附着于红细胞上，抗