临床输血学实验

,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,Testing Specimen Requirement,2ml EDTA whole blood,Fill in the tube label(s)with name,临床输血学实验,Transfusion Medicine,(blood transfusion laboratories),上海市第六人民医院,Shanghai Sixth People Hospital,输血科,Transfusion Department,History,Ancient times,敬畏血液-生命的源泉-饮血疗法,The period 1500-1800,治疗精神病 血液有毒 放血疗法-人体血液循环，为输血奠定基础-血液的携氧功能，合理治疗手段,The discovery of blood groups,血型之父-,Landsteiner,人类血型的发现，为安全输血提供重要保证。,而且，在遗传学、人类学、法医学、免疫学、部分疾病的发病机制探讨上也具有重要意义。,While blood cell antigens:,Red cell antigens,HLA antigens,Granulocytes,-specific antigens,Platelets（22）,Part,one,Red blood cell group,目的要求：,掌握,ABO,血型的鉴定,熟悉,Rh,血型的鉴定,ABO SYSTEM,Principle of procedure,红细胞上具有,A,抗原者为,A,型-,Red blood cells possess A blood group antigens,有,B,抗原者为,B,型-,Red blood cells possess B blood group antigens,A,和,B,抗原都没有者为,O,型-,Red blood cells lack A and B blood group antigens,A,和,B,抗原都有者为,AB,型-,Red blood cells possess A and B blood group antigens,Rh,SYSTEM,The most complex in the red cells antigen,Individuals 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 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40多个。,涉及临床主要是5个抗原：,C、c、D、E、e,及其相应的特异性抗体。,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%saline,pipetter,reagents:,The monoclonal antibody(,IgM,)include Anti-A,、Anti-B、Anti-D,5%Standard RBC-,Affirmagen,Methods,Solid phase adherence test,Tube test,Gel test,Manual,Semi-automated system,Automated system,PROCEDURE,Specimen collection and preparation,Red blood cell suspensions(4%RBC)can be prepared using the following combinations of saline and packed red blood cells:,Saline Volume-2ml,Packed RBC Volume-100ul,被检者,RBC,用生理盐水配成2-5%悬液待用,(2,ml 0.9%NS+100ul,压积红细胞）,TEST PROCEDURE-tube test,正定型,：,取小试管三只，分别用记号笔标记后加抗,A、,抗,B、,抗,D,血清各一滴，然后加受检者,RBC,悬液一滴。,blood grouping,Add one drop of Anti-A、Anti-B、Anti-D to three tubes separately,Add one drop of a 2-5%RBC suspension to three tubes separately,TEST PROCEDURE,反定型,：,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,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,将上述混匀后静置数分钟或3000,r/m,离心15,s,肉眼判断凝集度。,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.,Record the reaction strength on the paper.,4+,一个结实的大凝块，背景清楚。,3+,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。,1+,小凝块，背景较浑浊。,+,或W+,细胞有微小凝块，背景红色浑浊，通常用显微镜观察。,MF,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。,0(,negative),阴性，红细胞呈游离状态，无凝集象,H(,hemolysis,),溶血，有游离血红蛋白。,Evaluate the agglutination strength,Anti-A,Anti-B,Anti-D,Ac,Bc,Oc,grouping,+,+,+,A,RhD,positive,+,+,+,B,RhD,positive,+,+,+,O,RhD,positive,+,+,+,AB,RhD,positive,Interpretation of results,Grouping anomalies,Cold auto-antibodies,Acquired B,Unexpected mixed field reactions,Partial or weak D,D negative,Intrauterine transfusions,RhD,negative verification,Reagents:three different Anti-D human(,IgG,)monoclonal antibody,Method,Indirect,antiglobulin,test(IAT),Gene test,Indirect,antiglobulin,test,Incubating the donors red cells with the,Rh,typing sera,Washing the cells three times,Adding AHG,Centrifuging the mixture,Observing it for agglutination,O,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,O,A,B,AB,ABO group and amativeness,female,male,Part two,Red cell compatibility testing-,Crossmatch,目的要求：,掌握交叉配血试验的原理,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 not 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-,Major,crossmatch,(most important)-comparing donor,eryghrocytes,to recipient serum antibodies in recipient serum against donor erythrocytes.,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 cells