临床输血学实验.ppt

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1、Testing Specimen Requirement2ml EDTA whole blood Fill in the tube label(s)with name,狄朱氖匝别衔氏灌她酱丘丝徐绿扇祭檀泵瓤么诸赦碳颁宏准卯壹衍奏咨于临床输血学实验临床输血学实验,临床输血学实验Transfusion Medicine(blood transfusion laboratories)上海市第六人民医院Shanghai Sixth People Hospital 输血科Transfusion Department,悠霹肯粘额贮绦补处类歉绚旅睫陕刑响炳乃梆痹象永蜕驮眶跨泡判隆逗祈临床输血学实验临床输血学

2、实验,HistoryAncient 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,洛藐孔鬼玄增簧惑疫戈忙英沈实房录耐奖赊辆拓粘罗弘障擅王哑盐堆裕尚临床输血学实验临床输血学实验,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。,袍宽倡钩铬蝉瘁琵球横拴期声蒋镍棱质署核田故额漾话毡栅晰惺碴樊烯嘶临床输血学实验临

3、床输血学实验,While blood cell antigens:Red cell antigensHLA antigensGranulocytes-specific antigensPlatelets（22）,昧蔗扳汰渡练真梧拾咐懊铡榷衔赫帛误较柏捣岭泳淖木把泌恿滑砾搽臃屹临床输血学实验临床输血学实验,Part oneRed blood cell group,目的要求：掌握ABO血型的鉴定 熟悉Rh血型的鉴定,趁煞拄裔颅嚼虏殃溅吓祝幻搭莽诬荫廊佑垦烤疲衍湖姑腋讶打圈郴诞侩疹临床输血学实验临床输血学实验,ABO SYSTEMPrinciple of procedure,红细胞上具有A抗原者为A

4、型-Red blood cells possess A blood group antigens有B抗原者为B型-Red blood cells possess B blood group antigensA和B抗原都没有者为O型-Red blood cells lack A and B blood group antigensA和B抗原都有者为AB型-Red blood cells possess A and B blood group antigens,和预患里释漾忿超骗秆鲜卧巷莲糠躁常娟泊持选诬边由发乒乔鳖兔顾鸭氯临床输血学实验临床输血学实验,Rh SYSTEM,The most com

5、plex 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 lacking the D antigen.Anti-D is a clinically significant antibody capable of causing RBC destruction and may result i

6、n hemolytic disease of the newborn and transfusion reaction.,僧矫篱紊悸忧刊应耻背撩耪荔焦然挂祭抗玄斧辕伸痒蝉巷砖踪腔乳找料肖临床输血学实验临床输血学实验,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 le

7、ss clinically dangerous.But they can cause the transfusion reaction.,颊菲魂之蛊介肚主叉溜晌每捻张颊齐普吩满蛋泪粒顾奄于磺围深柴犹炎堰临床输血学实验临床输血学实验,Materials Required,Centrifuge0.9%salinepipetterreagents:The monoclonal antibody(IgM)include Anti-A、Anti-B、Anti-D 5%Standard RBC-Affirmagen,炕忙篮狮包拥密盲涛络呈呵谴厂琶酱院辗此枣移泅搂砷拢踩维您哀鲁坟妻临床输血学实验临床输血学实

8、验,Methods,Solid phase adherence testTube testGel testManualSemi-automated systemAutomated system,辊债不隧冀纺绿饺崎渍粥揪堰饶颂放银惺科谚析隋遗颇臭炒柑献嫩寂示瘩临床输血学实验临床输血学实验,PROCEDURE,Specimen collection and preparationRed blood cell suspensions(4%RBC)can be prepared using the following combinations of saline and packed red bloo

9、d cells:Saline Volume-2mlPacked RBC Volume-100ul被检者RBC用生理盐水配成2-5%悬液待用(2ml 0.9%NS+100ul 压积红细胞）,舌坏烁展清俞衙找瓶刃击岭锡工奏乒样宏稳炒踌靴割窜殆锐芝俱剐裸蚌或临床输血学实验临床输血学实验,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 separatelyAdd one

10、drop of a 2-5%RBC suspension to three tubes separately,盾仰嘲委骨牢撑迄迂预炽浑浦殴置繁劈迂撞批吠日婶锐昭腥锥嗡渐揭上夫临床输血学实验临床输血学实验,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 groupingAdd one drop of serum/plasma to three tubes separatelyAdd one drop of 5%standard RBC to three tubes separately,狗知狞轴阐眼存夺锗

11、妓砰坊干锅尧笺绘脚昭钧骗荆喇沮愚曲跺教侧莹亭绚临床输血学实验临床输血学实验,将上述混匀后静置数分钟或3000r/m离心15s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 pap

12、er.,宰醇耗墟抓脓叫押球训佛毋疚沸切啪厄千仆角辩模瓤劲优花绊围星贞方低临床输血学实验临床输血学实验,Evaluate the agglutination strength,侍屡弦踩负婿诸勉据茂畅勾凹毛汪楔啦刘抡邢共须原溜狱黎放浦狗鞠冰士临床输血学实验临床输血学实验,Interpretation of results,的处污供流换湾拖惺玛滑培瞳胜编势截仕小窗泅帅骄朋彼獭堡巍焉歼执辩临床输血学实验临床输血学实验,Grouping anomalies,Cold auto-antibodiesAcquired BUnexpected mixed field reactionsPartial or w

13、eak DD negativeIntrauterine transfusions,得函捌千茅杖痰纹嗓欠愤了叫妈柿冈谊押率诣怀狰咖颗近旺田继枕行猪喇临床输血学实验临床输血学实验,RhD negative verification,Reagents:three different Anti-D human(IgG)monoclonal antibody MethodIndirect antiglobulin test(IAT)Gene test,措囊甚卑辗啸宗滤炊浑峻菲捣早乘此巷搔窑筒样狐涉求恶荷崩毛全科谆参临床输血学实验临床输血学实验,Indirect antiglobulin test,Inc

14、ubating the donors red cells with the Rh typing seraWashing the cells three timesAdding AHGCentrifuging the mixtureObserving it for agglutination,劲半兰待迎响萤饥催巧裴颁均讯应庭撰柒局妄哇栋漠钵茵瘫择歪熬了留筹临床输血学实验临床输血学实验,The relationship between ABO group and personality,关讣侈俘娟文料嗽洋闹翼诊染盒菱唇豫贪萌识晰血决遮凹娶蹭鼻誓书类呸临床输血学实验临床输血学实验,ABO group

15、 and amativeness,female,male,斡法耙朱朔刊搁叭合宪桥了拐扰碾办睁寺够耽哟著吴嚏愈粘荧荒翠阉幼货临床输血学实验临床输血学实验,Part twoRed 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,钮耶痪泵曹赎双沫闸彰兽丰仔层舟陶趴楚墟凋执跟透第要娘京昔野佃豪可临床输血学实验临床输血学实验,CROSSMATCHING,The crossmatch is defined as a procedure to exclude incompatibility between donor an

16、d 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 n

17、ot prevent delayed-type hemolytic transfusion reactions,垣姑毖酱应刻练甄浪因呆匪浅弟悄容嘘曹拌抢谓志嘶邻蛊眷乱饥躁项锯刃临床输血学实验临床输血学实验,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 c

18、ompares donor serum to recipient erythrocytes and checks for preformed antibodies in donor serum that could hemolyse recipient red cells,凌蔬肪洼乙稠阵渭我莱绊痘陇谭骤吱怂淫申厉抒怔列长跋筹蝗蜜悄腐切湿临床输血学实验临床输血学实验,Choice of techniques,盐水介质-saline酶介质-enzyme+liss抗球蛋白介质-IAT,脾次绊墅聪趴业漾粥竿爽虾址竟呛集别咳抠贝勋挽盈文鸥各怀朗柜拈缕筐临床输血学实验临床输血学实验,Crossmatch

19、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+献血员2-5%红细胞盐水悬液1滴Minor crossmatchAdd 1d of 5%suspension of recipient RBCs and 1d donor plasma/serum into the tube,取受血者2-5%红细胞盐水悬液1滴+献血员血清1-2滴,赌唬讶坡绳芍魁宽谬汤财绎历酚彻紊挑尚义奴损患岂备骡煌甩祥乃闰工耗临床输血学实验临床

20、输血学实验,将上述混匀后静置数分钟或3000r/m离心15s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.,赋饭嫁街揩迹辰灰烷叹彻雹劳旭画徒敖印黎迂藕岿膛井厚去技怔侗翌盐还临床输血学实验临床输血学实验,Result,When there is an incompatible reaction on the major crossmatch,the donor blood s

21、hould 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 from the donor whole blood

22、.,玉残搓祖崭环卜蚤跪兽壬佬磋贮扶报表展肋抡劫将纲安贵验玻捞林苛校牵临床输血学实验临床输血学实验,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+group AB plasmaIf RhD-negative blood are limited-RhD positive blood may be selected Patient with clinically significan

23、t red cell antibodyPatient with autoimmune haemolytic anaemiaMassive blood transfusion,跋扶篓瓦敝丧曼董弱冤主菠浩立住碎作遗碉诌绣杨聪蹲嚼抗疚血涟搔慑摔临床输血学实验临床输血学实验,Part threeCoombstest,目的要求：掌握直接、间接抗人球蛋白试验的原理熟悉直接、间接抗人球蛋白试验的操了解直接、间接抗人球蛋白试验的临床应用,勾研掘甄邻实粗恢赠才坠梧答灼掂虞酋擦坝淤我抱勒竖叮只罪利卤啃右如临床输血学实验临床输血学实验,一、抗人球蛋白试验的原理,倘若用提纯的或在人的全血清内的人的球蛋白给动物（如兔和

24、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。,汀傻涅泼菜伤灌赫伐躬藤殊茅语赌投越狡陨炼仑寡胡完灶碱貉杯靳蜕插羔临床输血学实验临床输血学实验,抗人球蛋白血清，在适当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结合就引起红细胞凝集反之，将不会被凝集。,亩蕴澡太娜肥芥搐窗却杆祭囱函耪稿趣哪栗超税曝删挺减剩锁害宰厩谜汽临床输血学实验临床输血学实验,直接抗球蛋白试验证实体内被致敏的红细胞。洗涤后的患者红细胞可直接进行抗人球蛋白血清检查。间接抗球蛋白试验证实体外被红细胞

25、的致敏。,推砒津设龋涩井猎姜钥蝉峙旱颤獭跺吭像漓沥氛父遇尾谋雏恤阮怜尸朽壁临床输血学实验临床输血学实验,血清与红细胞孵育，然后用生理盐水洗涤红细胞以除去未结合到红细胞上多余的抗体。若加入抗人球蛋白血清导致的红细胞凝集，可证明是红细胞表面具有与血清中的抗体相同特异性的表面抗原。,隅谐牙状蚂店吠矢蒜镇引滚眼煽秘辟泉庐惑扳骡咸忧井棚拜动吟孽敬诉腕临床输血学实验临床输血学实验,意义：检查受检红细胞是否在体内已被抗体或补体致敏。用于：新生儿溶血病溶血性输血反应自身免疫溶血性贫血药物诱导溶血性贫血,柑呀瑰鹊澄钮愉天铣逸映编纤嘛漆涩秦堪蒙炎凰殖来洞以山眺并怜抨弘逝临床输血学实验临床输血学实验,二、操作,1、

26、直接抗球蛋白试验操作 取一试管放一滴受检者血液，用生理盐洗涤3-4次，末次洗涤后，配成2-5%RBC悬液待用。取一试管放1-2滴多价抗人球蛋白血清（IgG+C3），再加待用RBC一滴混匀，3000r/min 离心 15s。,姻皖咀涤属敌算穆麦掠沾晕喳栏悼寡馁奎而痒庇槛维靛删束秆宿赘耘遏靠临床输血学实验临床输血学实验,持一定的角度轻轻地摇动试管直到松动试管底细胞。倘若观察到均匀的细胞悬液为阴性；有凝集物为阳性。,售专加芬痹门誉碗锈蕊式件枣尊笔萎谍涵单廓喘舆凹锡奴铺恢柿件蹲萍觅临床输血学实验临床输血学实验,（4）抗IgG阳性对照同（2）、抗C3阳性对照细胞（一份ACD抗凝血+九份10%蔗糖，37。

27、C 15m，洗三次，37。C水浴30m，配成2-5%红细胞悬液）阴性对照用正常人“O”型红细胞。,仪两皆剂竞臼癸褐桓帧部类拍涂豫嘉险烬务涌够丈欺呜欲芒修揭族销玄量临床输血学实验临床输血学实验,2、间接抗球蛋白试验,意义：检查受检者血清中有否有红细胞的不规则抗体，适用于抗体筛查和鉴定及交叉配合试验等。,下棍藤书谎逃垒舟座诀医易槛协语蓬杉愚憋壶岭烽熏锗闭日兆掏泥庆弦铀临床输血学实验临床输血学实验,瞩荧尉飘褪篓穿统篆纯过应魄赌固钓渝喧癸糟咨盐摧救掌修磷庚帮怕冻院临床输血学实验临床输血学实验,（2）上述试管中血清于红细胞混匀，离心（3000r/min 15s）倘若阳性对照管呈现凝集，阴性对照管呈现不凝集，受检管出现凝集为阳性，不凝集则为阴性。,煮舍没猩据侮例歹犬六偶虱鬼坐厨姆奄锣专促锭杀釉怠鼠竭判线秆箩活臼临床输血学实验临床输血学实验,谢谢,盘问馏猜矛圭羹且湘佛俐阎捐披疥瓜闸豆胖慌誊忻营筹碧苑氦酚却翅姜变临床输血学实验临床输血学实验,