臨床輸血學(xué)實(shí)驗(yàn)講解課件

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

2、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The discovery of blood groups 血型之父-LandsteinerHistory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。人類血型的發(fā)現(xiàn)，為安全輸血提供重要保證。While blood cell antigens:Red cell antigensHLA antigensGranulocytes-specific antigensPlatelets（22）While blood cell an

3、tigens: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Part oneRed blood cell gro ABO SYSTEMPrinciple of procedure紅細(xì)胞上具有A抗原者為A型- Red blood cells possess A blood group antigens有B抗原者為B型- Red blood cells possess B blood group antigensA和B抗原都沒有者為O型- Red blood cells lack A and B blood group antigens

4、A和B抗原都有者為AB型- Red blood cells possess A and B blood group antigens ABO SYSTEMPrinciple of Rh SYSTEMThe most complex in the red cells antigenIndividuals who possess the D antigen are Rh positive, who lack it are Rh negative.The D antigen is capable of stimulating production of Anti-D in persons lacki

5、ng 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. Rh SYSTEMThe most complex in 已發(fā)現(xiàn)的抗原40多個(gè)。涉及臨床主要是5個(gè)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

6、 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.已發(fā)現(xiàn)的抗原40多個(gè)。Materials RequiredCentrifuge0.9% salinepipetterreagents:The monoclonal antibody(IgM) include Anti-A 、 Anti-B 、 Anti-D 5% St

7、andard RBC-AffirmagenMaterials RequiredCentrifugeMethodsSolid phase adherence testTube testGel testManualSemi-automated systemAutomated systemMethodsSolid phase adherence tPROCEDURESpecimen collection and preparationRed blood cell suspensions(4%RBC) can be prepared using the following combinations o

8、f 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）PROCEDURESpecimen collection aTEST PROCEDURE-tube test正定型： 取小試管三只，分別用記號(hào)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 separa

9、telyAdd one drop of a 2-5% RBC suspension to three tubes separatelyTEST PROCEDURE-tube test正定型：TEST PROCEDURE反定型： 取小試管三只，分別用記號(hào)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 groupingAdd one drop of serum/plasma to three tubes separatelyAdd one drop of 5% standard RBC to three tubes separatelyTEST PROCEDURE反定

10、型：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。Shake 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. 臨床輸血學(xué)實(shí)驗(yàn)講解課件4+一個(gè)結(jié)實(shí)的大凝塊，背景清楚。3+數(shù)個(gè)結(jié)

11、實(shí)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(xì)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(xì)胞形成較大的凝集塊，而周圍是游離的紅細(xì)胞。0(negative)陰性，紅細(xì)胞呈游離狀態(tài)，無凝集象H(hemolysis)溶血，有游離血紅蛋白。Evaluate the agglutination strength4+一個(gè)結(jié)實(shí)的大凝塊，背景清楚。3+數(shù)個(gè)結(jié)實(shí)的凝塊，背景清楚Anti-AAnti-BAnti-DAcBcOcgrouping+A RhD positive+B RhD positive+O RhD positive+AB R

12、hD positiveInterpretation of results Anti-AAnti-BAnti-DAcBcOcgroupiGrouping anomaliesCold auto-antibodiesAcquired BUnexpected mixed field reactionsPartial or weak DD negativeIntrauterine transfusionsGrouping anomaliesCold auto-anRhD negative verification Reagents: three different Anti-D human (IgG)

13、monoclonal antibody MethodIndirect antiglobulin test (IAT)Gene test RhD negative verification ReIndirect antiglobulin testIncubating the donors red cells with the Rh typing seraWashing the cells three timesAdding AHGCentrifuging the mixtureObserving it for agglutinationIndirect antiglobulin testIncu

14、O有一種表現(xiàn)自己的性格,行動(dòng)受感情左右,喜歡政治,好勝心強(qiáng),對(duì)立斗爭(zhēng)有戒心與喜歡交友并存,能看清形勢(shì),很注意別人對(duì)自己的好惡.有信念和理想.率直,具有現(xiàn)時(shí)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(duì)別人情緒及環(huán)境變化有敏銳反應(yīng),但對(duì)別人的想法缺乏客觀理解.思想行動(dòng)慎重而有規(guī)律,個(gè)性內(nèi)向,沒個(gè)性謹(jǐn)小慎微固步自封,遵從常理,重視家庭,富于計(jì)劃性.擅長(zhǎng)以說服和說明的調(diào)子講話B開放型對(duì)人缺乏戒心,情緒善變,活動(dòng)力強(qiáng),不原受約束,行動(dòng)奔放光明正大反復(fù)無常,富于靈活性,歡熱鬧場(chǎng)合.固執(zhí)己見,善于批評(píng),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(huì)上

15、施展能力,注意別人對(duì)自己的感情,厭惡表里不一.個(gè)性中庸,富有實(shí)踐精神計(jì)劃性強(qiáng)不喜歡指手劃腳容易疲勞.表情不豐富,正式場(chǎng)合講話呆板,回避感情激烈的講話或事件The relationship between ABO group and personality O有一種表現(xiàn)自己的性格,行動(dòng)受感情左右,喜歡政治,好勝心強(qiáng),OABAB ABO group and amativenessfemalemaleOABAB ABO group and amativePart twoRed cell compatibility testing-Crossmatch 目的要求： 掌握交叉配血試驗(yàn)的原理 熟悉交叉配血

16、試驗(yàn)的操作Part twoRed cell compatibil CROSSMATCHING The 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 typing is not available agai

17、nst 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 CROSSM Immediate spin crossmatch主側(cè)配血-Major crossmatch (most important)-comparing donor eryghrocytes to recip

18、ient serum antibodies in recipient serum against donor erythrocytes. 次側(cè)配血-Minor crossmatch This compares donor serum to recipient erythrocytes and checks for preformed antibodies in donor serum that could hemolyse recipient red cells Immediate spin crossmatch主側(cè)配血Choice of techniques鹽水介質(zhì)-saline酶介質(zhì)-en

19、zyme + liss抗球蛋白介質(zhì)-IATChoice of techniques鹽水介質(zhì)-sal Crossmatch procedure Major crossmatch: Add 1d of 5% suspension of donor RBCs and 1d recipient plasma/serum into the tube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Minor crossmatchAdd 1d of 5% suspension of recipient RBCs and 1d donor plasma/serum into the tube, 取受血者

20、2-5%紅細(xì)胞鹽水懸液1滴+獻(xiàn)血員血清1-2滴 Crossmatch procedu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 45-60 angles.Evaluate two tubes for agglutination under the microscope.將上述混勻后靜置數(shù)分鐘或3000r/m離心15s肉眼判斷凝集ResultWhen there is an incompatible reaction on the major crossmatch,

21、 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. However, if the donated serum is likely to contribute substantially to the plasma volume of the recipient, the serum should be removed fro

22、m the donor whole blood.ResultWhen there is an incompaSelection of bloodRed cell components-the same ABO and RhD group as the patient If ABO identical blood is not available-group O RBC + group AB plasmaIf RhD-negative blood are limited-RhD positive blood may be selected Patient with clinically sign

23、ificant red cell antibodyPatient with autoimmune haemolytic anaemiaMassive blood transfusionSelection of bloodRed cell comPart threeCoombstest目的要求：掌握直接、間接抗人球蛋白試驗(yàn)的原理熟悉直接、間接抗人球蛋白試驗(yàn)的操了解直接、間接抗人球蛋白試驗(yàn)的臨床應(yīng)用Part threeCoombstest目的要求 一、抗人球蛋白試驗(yàn)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(dòng)物（如兔和山羊）注射，動(dòng)物對(duì)外來的蛋白質(zhì)將會(huì)產(chǎn)生抗體，即抗人球蛋白。 一、抗人球蛋白試驗(yàn)的原理 倘若用提 抗人球蛋白血清，在適當(dāng)處理之后將會(huì)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(huì)與球蛋白結(jié)合。因?yàn)榍虻鞍赘街诩t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(huì)被凝集。 抗人球蛋白血清，在適當(dāng)處理之后將會(huì)與人球蛋白發(fā)生直接抗球