臨床輸血學(xué)實(shí)驗(yàn)

1、testing specimen requirement2ml edta whole blood fill in the tube label(s) with name 臨床輸血學(xué)實(shí)驗(yàn)臨床輸血學(xué)實(shí)驗(yàn)transfusion medicine(blood transfusion laboratories)上海市第六人民醫(yī)院上海市第六人民醫(yī)院shanghai sixth people hospital 輸血科輸血科transfusion departmenthistoryancient times 敬畏血液敬畏血液-生命的源泉生命的源泉-飲血療飲血療法法the period 1500-1800 治療

2、精神病治療精神病 血液有毒血液有毒 放血療法放血療法-人體血液循環(huán)，為輸血奠定基礎(chǔ)人體血液循環(huán)，為輸血奠定基礎(chǔ)-血液血液的攜氧功能，合理治療手段的攜氧功能，合理治療手段the discovery of blood groups 血型之父血型之父-landsteiner人類血型的發(fā)現(xiàn)，為安全輸人類血型的發(fā)現(xiàn)，為安全輸血提供重要保證。血提供重要保證。而且，在遺傳學(xué)、人類學(xué)、而且，在遺傳學(xué)、人類學(xué)、法醫(yī)學(xué)、免疫學(xué)、部分疾病法醫(yī)學(xué)、免疫學(xué)、部分疾病的發(fā)病機(jī)制探討上也具有重的發(fā)病機(jī)制探討上也具有重要意義。要意義。while blood cell antigens:red cell antigenshla

3、 antigensgranulocytes-specific antigensplatelets（22）part onered blood cell group目的要求：目的要求： 掌握掌握abo血型的鑒定血型的鑒定 熟悉熟悉rh血型的鑒定血型的鑒定 abo systemprinciple of procedure紅細(xì)胞上具有紅細(xì)胞上具有a抗原者為抗原者為a型型- red blood cells possess a blood group antigens有有b抗原者為抗原者為b型型- red blood cells possess b blood group antigensa和和b抗原都沒

4、有者為抗原都沒有者為o型型- red blood cells lack a and b blood group antigensa和和b抗原都有者為抗原都有者為ab型型- red blood cells possess a and b blood group antigensrh 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 s

5、timulating 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. 已發(fā)現(xiàn)的抗原已發(fā)現(xiàn)的抗原40多個(gè)。多個(gè)。涉及臨床主要是涉及臨床主要是5個(gè)抗原：個(gè)抗原：c、c、d、e、e及其相應(yīng)的特異性抗體。及其相應(yīng)的特異性抗體。four

6、 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 requiredcentrifuge0.9% salinepipetterreagents:the monoclonal antibody(igm)

7、 include anti-a 、 anti-b 、 anti-d 5% standard rbc-affirmagenmethodssolid phase adherence testtube testgel testmanualsemi-automated systemautomated systemprocedurespecimen collection and preparationred blood cell suspensions(4%rbc) can be prepared using the following combinations of saline and packed

8、 red blood cells:saline volume-2mlpacked rbc volume-100ul被檢者被檢者rbc用生理鹽水配成用生理鹽水配成2-5%懸液待用懸液待用(2ml 0.9% ns +100ul 壓積紅細(xì)胞）壓積紅細(xì)胞）test procedure-tube test正定型：正定型： 取小試管三只，分別用記號(hào)筆標(biāo)記后加抗取小試管三只，分別用記號(hào)筆標(biāo)記后加抗a a、抗抗b b、抗、抗d d血清各一滴，然后加受檢者血清各一滴，然后加受檢者rbcrbc懸液懸液一滴。一滴。blood grouping add one drop of anti-a、anti-b、anti-d

9、 to three tubes separatelyadd one drop of a 2-5% rbc suspension to three tubes separatelytest procedure反定型：反定型： 取小試管三只，分別用記號(hào)筆標(biāo)記后加受檢取小試管三只，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)者血清一滴，然后加標(biāo)準(zhǔn)a a、b b及及o o型型rbcrbc各一滴各一滴。reverse groupingreverse groupingadd one drop of serum/plasma to three tubes separatelyadd one drop of

10、 5% standard rbc to three tubes separately將上述混勻后靜置數(shù)分鐘或?qū)⑸鲜龌靹蚝箪o置數(shù)分鐘或3000r/m3000r/m離心離心15s15s肉眼判肉眼判斷凝集度。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.rec

11、ord the reaction strength on the paper. 4+一個(gè)結(jié)實(shí)的大凝塊，背景清楚。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。數(shù)個(gè)結(jié)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。中等大小的凝塊加小凝塊，背景清楚。1+小凝塊，背景較渾濁。小凝塊，背景較渾濁。+或或w+細(xì)胞有微小凝塊，背景紅色渾濁，通常細(xì)胞有微小凝塊，背景紅色渾濁，通常用顯微鏡觀察。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。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

12、集象陰性，紅細(xì)胞呈游離狀態(tài)，無凝集象evaluate the agglutination strengthevaluate the agglutination strengthanti-aanti-aanti-banti-banti-danti-dacacbcbcococgrouping+ + + +a rhd a rhd positive+ + + +b rhd b rhd positive+ + + +o rhd o rhd positive+ + + +ab rhd ab rhd positiveinterpretation of results grouping anomaliesco

13、ld auto-antibodiesacquired bunexpected mixed field reactionspartial or weak dd negativeintrauterine transfusionsrhd negative verification reagents: three different anti-d human (igg) monoclonal antibody methodindirect antiglobulin test (iat)gene test indirect antiglobulin testincubating the donors r

14、ed cells with the rh typing serawashing the cells three timesadding ahgcentrifuging the mixtureobserving it for agglutinationo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

15、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 oabab abo group and amativenessfemalemalepart t

16、wored cell compatibility testing-crossmatch 目的要求：目的要求： 掌握交叉配血試驗(yàn)的原理掌握交叉配血試驗(yàn)的原理 熟悉交叉配血試驗(yàn)的操作熟悉交叉配血試驗(yàn)的操作 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 n

17、ot be evident on blood typing (as blood typing 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)配血主側(cè)配血-major

18、crossmatch (most important)-comparing donor eryghrocytes to recipient serum antibodies in recipient serum against donor erythrocytes. 次側(cè)配血次側(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 cel

19、lschoice of techniques鹽水介質(zhì)鹽水介質(zhì)-saline酶介質(zhì)酶介質(zhì)-enzyme + liss抗球蛋白介質(zhì)抗球蛋白介質(zhì)-iat crossmatch procedure major crossmatch: add 1d of 5% suspension of donor rbcs and 1d recipient plasma/serum into the tube取受血者血清取受血者血清1-2滴滴+獻(xiàn)血員獻(xiàn)血員2-5%紅細(xì)胞鹽水懸紅細(xì)胞鹽水懸液液1滴滴minor crossmatchadd 1d of 5% suspension of recipient rbcs and

20、 1d donor plasma/serum into the tube, 取受血者取受血者2-5%紅細(xì)胞鹽水懸液紅細(xì)胞鹽水懸液1滴滴+獻(xiàn)血員血獻(xiàn)血員血清清1-2滴滴將上述混勻后靜置數(shù)分鐘或?qū)⑸鲜龌靹蚝箪o置數(shù)分鐘或3000r/m3000r/m離心離心15s15s肉眼判斷凝集度。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.resultwhen there is an inc

21、ompatible reaction 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. however, if the donated serum is likely to contribute substantially to the plasma volume of the

22、 recipient, the serum should be removed from the donor whole blood.selection 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 cl

23、inically significant red cell antibodypatient with autoimmune haemolytic anaemiamassive blood transfusionpart threecoombstest目的要求：目的要求：掌握直接、間接抗人球蛋白試驗(yàn)掌握直接、間接抗人球蛋白試驗(yàn)的原理的原理熟悉直接、間接抗人球蛋白試驗(yàn)熟悉直接、間接抗人球蛋白試驗(yàn)的操的操了解直接、間接抗人球蛋白試驗(yàn)了解直接、間接抗人球蛋白試驗(yàn)的臨床應(yīng)用的臨床應(yīng)用 一、抗人球蛋白試驗(yàn)的原理一、抗人球蛋白試驗(yàn)的原理 倘若用提純的或在人的全血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

24、（如兔清內(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，即抗人球蛋白質(zhì)將會(huì)產(chǎn)生抗體，即抗人球蛋白。白。 抗人球蛋白血清，在適當(dāng)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)，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，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，將結(jié)合就引起紅細(xì)胞凝集反之，將不會(huì)被凝

25、集。不會(huì)被凝集。 直接抗球蛋白試驗(yàn)直接抗球蛋白試驗(yàn)證實(shí)體內(nèi)被致敏的紅細(xì)胞。證實(shí)體內(nèi)被致敏的紅細(xì)胞。洗滌后的患者紅細(xì)胞可直接洗滌后的患者紅細(xì)胞可直接進(jìn)行抗人球蛋白血清檢查進(jìn)行抗人球蛋白血清檢查。間接抗球蛋白試驗(yàn)間接抗球蛋白試驗(yàn)證實(shí)體外被紅細(xì)胞的致敏證實(shí)體外被紅細(xì)胞的致敏。血清與紅細(xì)胞孵育，然血清與紅細(xì)胞孵育，然后用生理鹽水洗滌紅細(xì)后用生理鹽水洗滌紅細(xì)胞以除去未結(jié)合到紅細(xì)胞以除去未結(jié)合到紅細(xì)胞上多余的抗體。若加胞上多余的抗體。若加入抗人球蛋白血清導(dǎo)致入抗人球蛋白血清導(dǎo)致的紅細(xì)胞凝集，可證明的紅細(xì)胞凝集，可證明是紅細(xì)胞表面具有與血是紅細(xì)胞表面具有與血清中的抗體相同特異性清中的抗體相同特異性的表面抗原。的表面抗原。意義：意義：檢查受檢紅細(xì)胞是否在體內(nèi)已檢查受檢紅細(xì)胞是否在體內(nèi)已被抗體或補(bǔ)體致敏。被抗體或補(bǔ)體致敏。 用于：用于：新生兒溶血病新生兒溶血病溶血性輸血反應(yīng)溶血性輸血反應(yīng)自身免疫溶血性貧血自身免疫溶血性貧血藥物誘導(dǎo)溶血性貧血藥物誘導(dǎo)溶血性貧血二、操作二、操作 1、直接抗球蛋白試驗(yàn)操作、直接抗球蛋白試驗(yàn)操作 取一試管放一滴受檢者血液，用生取一試管放一滴受檢者血液，用生理鹽洗滌理鹽洗滌3-