临床输血学实验

1、testing specimen requirement2ml edta whole blood fill in the tube label(s) with name 临床输血学实验临床输血学实验transfusion medicine(blood transfusion laboratories)上海市第六人民医院上海市第六人民医院shanghai sixth people hospital 输血科输血科transfusion departmenthistoryancient times 敬畏血液敬畏血液-生命的源泉生命的源泉-饮血疗饮血疗法法the period 1500-1800 治疗

2、精神病治疗精神病 血液有毒血液有毒 放血疗法放血疗法-人体血液循环，为输血奠定基础人体血液循环，为输血奠定基础-血液血液的携氧功能，合理治疗手段的携氧功能，合理治疗手段the discovery of blood groups 血型之父血型之父-landsteiner人类血型的发现，为安全输人类血型的发现，为安全输血提供重要保证。血提供重要保证。而且，在遗传学、人类学、而且，在遗传学、人类学、法医学、免疫学、部分疾病法医学、免疫学、部分疾病的发病机制探讨上也具有重的发病机制探讨上也具有重要意义。要意义。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血型的鉴定血型的鉴定 熟悉熟悉rh血型的鉴定血型的鉴定 abo systemprinciple of procedure红细胞上具有红细胞上具有a抗原者为抗原者为a型型- red blood cells possess a blood group antigens有有b抗原者为抗原者为b型型- red blood cells possess b blood group antigensa和和b抗原都没

4、有者为抗原都没有者为o型型- red blood cells lack a and b blood group antigensa和和b抗原都有者为抗原都有者为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. 已发现的抗原已发现的抗原40多个。多个。涉及临床主要是涉及临床主要是5个抗原：个抗原：c、c、d、e、e及其相应的特异性抗体。及其相应的特异性抗体。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被检者被检者rbc用生理盐水配成用生理盐水配成2-5%悬液待用悬液待用(2ml 0.9% ns +100ul 压积红细胞）压积红细胞）test procedure-tube test正定型：正定型： 取小试管三只，分别用记号笔标记后加抗取小试管三只，分别用记号笔标记后加抗a a、抗抗b b、抗、抗d d血清各一滴，然后加受检者血清各一滴，然后加受检者rbcrbc悬液悬液一滴。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： 取小试管三只，分别用记号笔标记后加受检取小试管三只，分别用记号笔标记后加受检者血清一滴，然后加标准者血清一滴，然后加标准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将上述混匀后静置数分钟或将上述混匀后静置数分钟或3000r/m3000r/m离心离心15s15s肉眼判肉眼判断凝集度。断凝集度。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+一个结实的大凝块，背景清楚。一个结实的大凝块，背景清楚。3+数个结实的凝块，背景清楚。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。中等大小的凝块加小凝块，背景清楚。1+小凝块，背景较浑浊。小凝块，背景较浑浊。+或或w+细胞有微小凝块，背景红色浑浊，通常细胞有微小凝块，背景红色浑浊，通常用显微镜观察。用显微镜观察。mf混合外观凝集，少数红细胞形成较大的混合外观凝集，少数红细胞形成较大的凝集块，而周围是游离的红细胞。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

12、集象阴性，红细胞呈游离状态，无凝集象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

15、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： 掌握交叉配血试验的原理掌握交叉配血试验的原理 熟悉交叉配血试验的操作熟悉交叉配血试验的操作 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主侧配血主侧配血-major

18、crossmatch (most important)-comparing donor eryghrocytes to recipient serum antibodies in recipient serum against donor erythrocytes. 次侧配血次侧配血-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盐水介质盐水介质-saline酶介质酶介质-enzyme + liss抗球蛋白介质抗球蛋白介质-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+献血员献血员2-5%红细胞盐水悬红细胞盐水悬液液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%红细胞盐水悬液红细胞盐水悬液1滴滴+献血员血献血员血清清1-2滴滴将上述混匀后静置数分钟或将上述混匀后静置数分钟或3000r/m3000r/m离心离心15s15s肉眼判断凝集度。肉眼判断凝集度。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、间接抗人球蛋白试验掌握直接、间接抗人球蛋白试验的原理的原理熟悉直接、间接抗人球蛋白试验熟悉直接、间接抗人球蛋白试验的操的操了解直接、间接抗人球蛋白试验了解直接、间接抗人球蛋白试验的临床应用的临床应用 一、抗人球蛋白试验的原理一、抗人球蛋白试验的原理 倘若用提纯的或在人的全血倘若用提纯的或在人的全血清内的人的球蛋白给动物

24、（如兔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，即抗人球蛋白质将会产生抗体，即抗人球蛋白。白。 抗人球蛋白血清，在适当抗人球蛋白血清，在适当处理之后将会与人球蛋白发生特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，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合。因为球蛋白附着于红细胞上，抗球蛋白血清与球蛋白分子的，抗球蛋白血清与球蛋白分子的结合就引起红细胞凝集反之，将结合就引起红细胞凝集反之，将不会被凝

25、集。不会被凝集。 直接抗球蛋白试验直接抗球蛋白试验证实体内被致敏的红细胞。证实体内被致敏的红细胞。洗涤后的患者红细胞可直接洗涤后的患者红细胞可直接进行抗人球蛋白血清检查进行抗人球蛋白血清检查。间接抗球蛋白试验间接抗球蛋白试验证实体外被红细胞的致敏证实体外被红细胞的致敏。血清与红细胞孵育，然血清与红细胞孵育，然后用生理盐水洗涤红细后用生理盐水洗涤红细胞以除去未结合到红细胞以除去未结合到红细胞上多余的抗体。若加胞上多余的抗体。若加入抗人球蛋白血清导致入抗人球蛋白血清导致的红细胞凝集，可证明的红细胞凝集，可证明是红细胞表面具有与血是红细胞表面具有与血清中的抗体相同特异性清中的抗体相同特异性的表面抗原。的表面抗原。意义：意义：检查受检红细胞是否在体内已检查受检红细胞是否在体内已被抗体或补体致敏。被抗体或补体致敏。 用于：用于：新生儿溶血病新生儿溶血病溶血性输血反应溶血性输血反应自身免疫溶血性贫血自身免疫溶血性贫血药物诱导溶血性贫血药物诱导溶血性贫血二、操作二、操作 1、直接抗球蛋白试验操作、直接抗球蛋白试验操作 取一试管放一滴受检者血液，用生取一试管放一滴受检者血液，用生理盐洗涤理盐洗涤3-