TestingSpecimenRequirement2mlEDTAwholebloodFillinthetubelabel(s)withname

1精品醫(yī)學(xué)臨床輸血學(xué)實驗

TransfusionMedicine(bloodtransfusionlaboratories)上海市第六人民醫(yī)院ShanghaiSixthPeopleHospital輸血科TransfusionDepartment2精品醫(yī)學(xué)HistoryAncienttimes敬畏血液-生命的源泉-飲血療法Theperiod1500-1800治療精神病血液有毒放血療法-人體血液循環(huán)，為輸血奠定基礎(chǔ)-血液的攜氧功能，合理治療手段Thediscoveryofbloodgroups血型之父--Landsteiner3精品醫(yī)學(xué)人類血型的發(fā)現(xiàn)，為安全輸血提供重要保證。而且，在遺傳學(xué)、人類學(xué)、法醫(yī)學(xué)、免疫學(xué)、部分疾病的發(fā)病機制探討上也具有重要意義。4精品醫(yī)學(xué)Whilebloodcellantigens:RedcellantigensHLAantigensGranulocytes-specificantigensPlatelets（22）5精品醫(yī)學(xué)

Partone

Redbloodcellgroup

目的要求：

掌握ABO血型的鑒定熟悉Rh血型的鑒定6精品醫(yī)學(xué)

ABOSYSTEM

Principleofprocedure

紅細胞上具有A抗原者為A型-RedbloodcellspossessAbloodgroupantigens有B抗原者為B型-RedbloodcellspossessBbloodgroupantigensA和B抗原都沒有者為O型-RedbloodcellslackAandBbloodgroupantigensA和B抗原都有者為AB型-RedbloodcellspossessAandBbloodgroupantigens7精品醫(yī)學(xué)RhSYSTEMThemostcomplexintheredcellsantigenIndividualswhopossesstheDantigenareRhpositive,wholackitareRhnegative.TheDantigeniscapableofstimulatingproductionofAnti-DinpersonslackingtheDantigen.Anti-DisaclinicallysignificantantibodycapableofcausingRBCdestructionandmayresultinhemolyticdiseaseofthenewbornandtransfusionreaction.8精品醫(yī)學(xué)已發(fā)現(xiàn)的抗原40多個。涉及臨床主要是5個抗原：C、c、D、E、e及其相應(yīng)的特異性抗體。Fourotherantigens(C、c、E、e)accountforalmostalloftheRh-relatedtransfusionproblems.TheyarelessantigenicthanD,and/ortheantibodiesarelessclinicallydangerous.Buttheycancausethetransfusionreaction.9精品醫(yī)學(xué)MaterialsRequiredCentrifuge0.9%salinepipetterreagents:Themonoclonalantibody(IgM)includeAnti-A

、Anti-B、Anti-D5%StandardRBC-Affirmagen10精品醫(yī)學(xué)MethodsSolidphaseadherencetestTubetestGeltestManualSemi-automatedsystemAutomatedsystem11精品醫(yī)學(xué)PROCEDURESpecimencollectionandpreparationRedbloodcellsuspensions(4%RBC)canbepreparedusingthefollowingcombinationsofsalineandpackedredbloodcells:SalineVolume---2mlPackedRBCVolume---100ul被檢者RBC用生理鹽水配成2-5%懸液待用(2ml0.9%NS+100ul壓積紅細胞）12精品醫(yī)學(xué)TESTPROCEDURE---tubetest正定型：取小試管三只，分別用記號筆標(biāo)記后加抗A、抗B、抗D血清各一滴，然后加受檢者RBC懸液一滴。bloodgroupingAddonedropofAnti-A、Anti-B、Anti-DtothreetubesseparatelyAddonedropofa2-5%RBCsuspensiontothreetubesseparately13精品醫(yī)學(xué)TESTPROCEDURE反定型：取小試管三只，分別用記號筆標(biāo)記后加受檢者血清一滴，然后加標(biāo)準A、B及O型RBC各一滴。reversegroupingAddonedropofserum/plasmatothreetubesseparatelyAddonedropof5%standardRBCtothreetubesseparately14精品醫(yī)學(xué)

將上述混勻后靜置數(shù)分鐘或3000r/m離心15s肉眼判斷凝集度。Shakegentlytomixcontentsofreactiontubes.Centrifugesixtubeswith3000r/s,15m.Shakegentlywith45-60angles.Evaluatesixtubesforagglutinationand/orhemolysisupontestcompletion.Recordthereactionstrengthonthepaper.

15精品醫(yī)學(xué)4+一個結(jié)實的大凝塊，背景清楚。3+數(shù)個結(jié)實的凝塊，背景清楚。2+中等大小的凝塊加小凝塊，背景清楚。1+小凝塊，背景較渾濁。+或W+細胞有微小凝塊，背景紅色渾濁，通常用顯微鏡觀察。MF混合外觀凝集，少數(shù)紅細胞形成較大的凝集塊，而周圍是游離的紅細胞。0(negative)陰性，紅細胞呈游離狀態(tài)，無凝集象H(hemolysis)溶血，有游離血紅蛋白。Evaluatetheagglutinationstrength16精品醫(yī)學(xué)Anti-AAnti-BAnti-DAcBcOcgrouping+++———ARhDpositive—+++——BRhDpositive——+++—ORhDpositive+++———ABRhDpositiveInterpretationofresults17精品醫(yī)學(xué)GroupinganomaliesColdauto-antibodiesAcquiredBUnexpectedmixedfieldreactionsPartialorweakDDnegativeIntrauterinetransfusions18精品醫(yī)學(xué)RhDnegativeverification

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

19精品醫(yī)學(xué)IndirectantiglobulintestIncubatingthedonor’sredcellswiththeRhtypingseraWashingthecellsthreetimesAddingAHGCentrifugingthemixtureObservingitforagglutination20精品醫(yī)學(xué)O有一種表現(xiàn)自己的性格,行動受感情左右,喜歡政治,好勝心強,對立斗爭有戒心與喜歡交友并存,能看清形勢,很注意別人對自己的好惡.有信念和理想.率直,具有現(xiàn)時與羅曼蒂克雙重性.言行帶有夸張性，講話帶有教育人和教訓(xùn)人的語氣A內(nèi)心想法不愿為人所知.對別人情緒及環(huán)境變化有敏銳反應(yīng),但對別人的想法缺乏客觀理解.思想行動慎重而有規(guī)律,個性內(nèi)向,沒個性謹小慎微固步自封,遵從常理,重視家庭,富于計劃性.擅長以說服和說明的調(diào)子講話B開放型對人缺乏戒心,情緒善變,活動力強,不原受約束,行動奔放光明正大反復(fù)無常,富于靈活性,歡熱鬧場合.固執(zhí)己見,善于批評,先下結(jié)論后作說明AB善于待人接物,很注意合情合理,期望在社會上施展能力,注意別人對自己的感情,厭惡表里不一.個性中庸,富有實踐精神計劃性強不喜歡指手劃腳容易疲勞.表情不豐富,正式場合講話呆板,回避感情激烈的講話或事件TherelationshipbetweenABOgroupandpersonality21精品醫(yī)學(xué)OABAB

ABOgroupandamativenessfemalemale22精品醫(yī)學(xué)Parttwo

Redcellcompatibilitytesting-Crossmatch

目的要求：掌握交叉配血試驗的原理熟悉交叉配血試驗的操作23精品醫(yī)學(xué)

CROSSMATCHING

Thecrossmatchisdefinedasaproceduretoexcludeincompatibilitybetweendonorandrecipient.Crossmatchingwillpickupincompatibilitiesbetweenthedonorandrecipientthatwillnotbeevidentonbloodtyping(asbloodtypingisnotavailableagainsteverybloodgroup,justthemajorones).Inaddition,thecrossmatchprocedurewillnotpickuplowtiterantibodiesandthuswillnotpreventdelayed-typehemolytictransfusionreactions24精品醫(yī)學(xué)

Immediatespincrossmatch主側(cè)配血---Majorcrossmatch(mostimportant)---comparingdonoreryghrocytestorecipientserumantibodiesinrecipientserumagainstdonorerythrocytes.次側(cè)配血---Minorcrossmatch

Thiscomparesdonorserumtorecipienterythrocytesandchecksforpreformedantibodiesindonorserumthatcouldhemolyserecipientredcells25精品醫(yī)學(xué)Choiceoftechniques鹽水介質(zhì)---saline酶介質(zhì)---enzyme+liss抗球蛋白介質(zhì)---IAT26精品醫(yī)學(xué)

Crossmatchprocedure

Majorcrossmatch:Add1dof5%suspensionofdonorRBCsand1drecipientplasma/serumintothetube取受血者血清1-2滴+獻血員2-5%紅細胞鹽水懸液1滴MinorcrossmatchAdd1dof5%suspensionofrecipientRBCsand1ddonorplasma/serumintothetube,取受血者2-5%紅細胞鹽水懸液1滴+獻血員血清1-2滴27精品醫(yī)學(xué)將上述混勻后靜置數(shù)分鐘或3000r/m離心15s肉眼判斷凝集度。Centrifugesixtubeswith3000r/s,15m.Shakesoftlywith45-60angles.Evaluatetwotubesforagglutinationunderthemicroscope.28精品醫(yī)學(xué)ResultWhenthereisanincompatiblereactiononthemajorcrossmatch,thedonorbloodshouldnotbetransfusionunderanycircumstances.Whenthereisanincompatiblereactionontheminorcrossmatch,thetransfusioncangoahead.However,ifthedonatedserumislikelytocontributesubstantiallytotheplasmavolumeoftherecipient,theserumshouldberemovedfromthedonorwholeblood.29精品醫(yī)學(xué)SelectionofbloodRedcellcomponents---thesameABOandRhDgroupasthepatientIfABOidenticalbloodisnotavailable---groupORBC+groupABplasmaIfRhD-negativebloodarelimited---RhDpositivebloodmaybeselectedPatientwithclinicallysignificantredcellantibodyPatientwithautoimmunehaemolyticanaemiaMassivebloodtransfusion30精品醫(yī)學(xué)

Partthree

Coombs’test

目的要求：掌握直接、間接抗人球蛋白試驗的原理熟悉直接、間接抗人球蛋白試驗的操了解直接、間接抗人球蛋白試驗的臨床應(yīng)用31精品醫(yī)學(xué)

一、抗人球蛋白試驗的原理

倘若用提純的或在人的全血清內(nèi)的人的球蛋白給動物（如兔和山羊）注射，動物對外來的蛋白質(zhì)將會產(chǎn)生抗體，即抗人球蛋白。32精品醫(yī)學(xué)抗人球蛋白血清，在適當(dāng)處理之后將會與人球蛋白發(fā)生特異性反應(yīng)。如果球蛋白分子，或者抗體或補體附著于紅細胞膜上，抗球蛋白血清將會與球蛋白結(jié)合。因為球蛋白附著于紅細胞上，抗