臨床輸血學(xué)實驗課件

1、Testing Specimen Requirement2ml EDTA whole blood Fill in the tube label(s) with name臨床輸血學(xué)實驗Transfusion Medicine(blood transfusion laboratories)上海市第六人民醫(yī)院Shanghai Sixth People Hospital輸血科Transfusion DepartmentHistoryAncient times 敬畏血液-生命的源泉-飲血療法The period 1500-1800 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

2、，合理治療手段The discovery of blood groups 血型之父-Landsteiner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(jī)制探討上也具有重要意義。While blood cell antigens:?Red cell antigens?HLA antigens?Granulocytes-specific antigensPlatelets（22）Part oneRed blood cell group目的要求：掌握ABO血型的鑒定熟悉Rh血型的鑒定ABO SYSTEMPrinciple of procedure?紅

3、細(xì)胞上具有A抗原者為A型-RedbloodcellspossessAbloodgroupantigens?有B抗原者為B型-RedbloodcellspossessBbloodgroupantigens?A和B抗原都沒有者為O型-RedbloodcellslackAandBbloodgroupantigens?A和B抗原都有者為AB型-RedbloodcellspossessAandBbloodgroupantigensRh SYSTEM?The most complex in the red cells antigen?Individuals who possess the D antige

4、n are Rh positive, who lack it are Rh negative.?The D antigen is capable of stimulating production of Anti-D in persons lacking the D antigen. ?Anti-D is a clinically significant antibody capable of causing RBC destruction and may result in hemolytic disease of the newborn and transfusion reaction.

5、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。Four other antigens(C、c、E、e) account for almost all of the Rh-related transfusion problems. They are less antigenic than D, and/or the antibodies are less clinically dangerous.But they can cause the transfusion reaction.Materials Required?Centrifuge?0.9% sa

6、line?pipetter?reagents:The monoclonal antibody(IgM) include Anti-A 、Anti-B 、Anti-D 5% Standard RBC-AffirmagenMethods?Solid phase adherence test?Tube test?Gel test?Manual?Semi-automated system?Automated systemPROCEDURESpecimen collection and preparationRed blood cell suspensions(4%RBC) can be prepare

7、d using the following combinations of saline and packed red blood cells:Saline Volume-2mlPacked RBC Volume-100ul被檢者RBC用生理鹽水配成2-5%懸液待用(2ml 0.9% NS +100ul 壓積紅細(xì)胞）TEST PROCEDURE-tube test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。blood grouping ?Add one drop of Anti-A、Anti-B、Anti-D to three tubes

8、separately?Add one drop of a 2-5% RBC suspension to three tubes separatelyTEST PROCEDURE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(zhǔn)A、B及O型RBC各一滴。reverse grouping?Add one drop of serum/plasma to three tubes separately?Add one drop of 5% standard RBC to three tubes separately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。?Sha

9、ke gently to mix contents of reaction tubes.?Centrifuge six tubes with 3000r/s,15m.?Shake gently with 45-60 angles.?Evaluate six tubes for agglutination and/or hemolysis upon test completion.?Record the reaction strength on the paper.Evaluate the agglutination strength4+3+2+1+或W+一個結(jié)實的大凝塊，背景清楚。數(shù)個結(jié)實的凝

10、塊，背景清楚。中等大小的凝塊加小凝塊，背景清楚。小凝塊，背景較渾濁。細(xì)胞有微小凝塊，背景紅色渾濁，通常用顯微鏡觀察?；旌贤庥^凝集，少數(shù)紅細(xì)胞形成較大的凝集塊，而周圍是游離的紅細(xì)胞。陰性，紅細(xì)胞呈游離狀態(tài)，無凝集象溶血，有游離血紅蛋白。MF0(negative)H(hemolysis)Interpretation of results Anti-AAnti-BAnti-DAcBcOcgrouping+A RhD positiveB RhD positiveO RhD positiveAB RhD positiveGrouping anomalies?Cold auto-antibodies?Ac

11、quired B?Unexpected mixed field reactions?Partial or weak D?D negative?Intrauterine transfusionsRhD negative verificationReagents: three different Anti-D human (IgG) monoclonal antibodyMethod?Indirect antiglobulin test (IAT)?Gene testIndirect antiglobulin test?Incubating the donors red cells with th

12、e Rh typing sera?Washing the cells three times?Adding AHG?Centrifuging the mixture?Observing it for agglutinationThe relationship between ABO group and personality 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(qiáng),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),

13、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(jǐn)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)子講話開放型對人缺乏戒心,情緒善變,活動力強(qiáng),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BAB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(qiáng)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ABO group and amativenessfemaleOBAABmalePart twoRed cell co

14、mpatibility testing-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CROSSMATCHINGThe crossmatch is defined as a procedure to exclude incompatibility between donor and recipient. Crossmatching will pick up incompatibilities between the donor and recipient that will not be evident on blood typing (as blood typin

15、g is not available against every blood group, just the major ones). In addition, the crossmatch procedure will not pick up low titer antibodies and thus will not prevent delayed-type hemolytic transfusion reactions Immediate spin crossmatch?主側(cè)配血-Majorcrossmatch(mostimportant)-comparingdonoreryghrocy

16、testorecipientserumantibodiesinrecipientserumagainstdonorerythrocytes.?次側(cè)配血-MinorcrossmatchThiscomparesdonorserumtorecipienterythrocytesandchecksforpreformedantibodiesindonorserumthatcouldhemolyserecipientredcellsChoice of techniques?鹽水介質(zhì)-saline?酶介質(zhì)-enzyme+liss?抗球蛋白介質(zhì)-IATCrossmatch procedureMajorcro

17、ssmatch:Add1dof5%suspensionofdonorRBCsand1drecipientplasma/serumintothetube取受血者血清1-2滴+獻(xiàn)血員2-5%紅細(xì)胞鹽水懸液1滴MinorcrossmatchAdd1dof5%suspensionofrecipientRBCsand1ddonorplasma/serumintothetube,取受血者2-5%紅細(xì)胞鹽水懸液1滴+獻(xiàn)血員血清1-2滴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。Centrifuge six tubes with 3000r/s,15m.Shake softly with

18、45-60 angles.Evaluate two tubes foragglutination under the microscope.Result?When there is an incompatiblereaction on the major crossmatch, the donor blood should not be transfusion under any circumstances. ?When there is an incompatible reaction on the minor crossmatch, the transfusion can go ahead

19、. However, if the donated serum is likely to contribute substantially to the plasma volume of the recipient, the serum should be removed from the donor whole blood.Selection of blood?Red cell components-the same ABO and RhD group as the patient ?If ABO identical blood is not available-group O RBC +

20、group AB plasma?If RhD-negative blood are limited-RhD positive blood may be selected ?Patient with clinically significant red cell antibody?Patient with autoimmune haemolytic anaemia?Massive blood transfusionPart threeCoombstest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?？谷饲虻鞍籽?，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(bǔ)體附著于紅細(xì)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(xì)胞上，抗球蛋白血清與球蛋白分子的結(jié)合就引起紅細(xì)胞凝集反之，將不會被凝集。直接抗球蛋白試驗證實體內(nèi)被致敏的紅細(xì)胞。洗滌后的患者紅細(xì)胞可直接進(jìn)行