抗-CD47单抗对输血相容性检测的干扰及处理

目的 评估抗-CD47单克隆抗体对输血相容性检测的干扰，研究去干扰方法，并对受试者输注效果进行评价。方法 收集本院参与接受天境和信达公司生产的抗-CD47单克隆抗体药物治疗的8名临床试验受试者标本，使用缺乏IgG4的抗人球蛋白试剂Gamma-clone、ZZAP试剂和药物独特型中和试剂等进行血型检测、意外抗体筛查、直接抗人球蛋白试验和交叉配血试验。结果 使用抗-CD47单抗治疗后，8名受试者中有5名ABO血型检定受到干扰；所有受试者直接抗人球蛋白凝集强度呈现2+～4+,且所有受试者意外抗体筛查结果都呈3+～4+。采用缺乏IgG4的抗人球蛋白试剂Gamma-clone的试管法进行意外抗体筛查试验，结果均呈现阴性，交叉配血试验均相合。采用药物独特型中和抗体进行意外抗体筛选，试验结果显示均为阴性，交叉配血试验均相合。使用CD47单抗药物导致贫血的患者均输注2U悬浮红细胞，评价经抗干扰处理后配血相合患者的输血效果，结果显示患者输血有效。结论 受试者使用CD47单抗药物后会干扰输血相容性检测，使用缺乏IgG4的抗人球蛋白试剂和药物独特型中和抗体可去除抗-CD47干扰，受试者输血效果良好。     

抗-CD47单克隆抗体干扰输血相容性检测1例

目的 探讨抗-CD47单抗对输血相容性检测的影响及处理措施。方法 对1名具有抗-CD47单抗用药史的患者血液标本进行输血相容性检测，分析抗-CD47单抗效价，鉴定巯基试剂对抗-CD47单抗的洗脱效果，利用中和抗体及抗球蛋白筛选与患者相配合的血液成分。结果 患者血型鉴定为A型，RhD(+),不规则抗体筛查、DAT试验、交叉配血均为3+或4+。利用中和抗体或Gamma-clone抗球蛋白均可为患者筛选到主侧配血相合的红细胞。患者输注后无不良反应，贫血状况得到明显改善。结论 利用中和抗体或Gamma-clone抗球蛋白处理患者血液标本，可以消除抗-CD47单抗对输血相容性检测的影响，保障临床用血的安全性、及时性和有效性。     

抗-CD38单克隆抗体对输血前检测试验的干扰及其处理措施

目的探讨抗-CD38单克隆抗体对输血前检测实验的干扰及其处理措施。方法收集2名接受抗-CD38单克隆抗体治疗的多发性骨髓瘤患者血样,分别进行ABO和Rh血型抗原定型、直接抗人球蛋白实验、不规则抗体筛选和鉴定、交叉配血实验;将不规则抗体筛选用试剂红细胞、抗体鉴定谱细胞、交叉配血用的献血者红细胞、平行对照用的O型K(+)E(+)红细胞与0.2 mol/L的DTT按照红细胞:DTT为1∶4的比例进行混合后,放入37℃水浴箱水浴30 min;再用上述DTT处理后的红细胞分别对患者血浆进行不规则抗体筛选实验、抗体鉴定实验和交叉配血实验。结果 2名患者的ABO和Rh血型均为O,CCDee;直接抗人球蛋白实验分别为阴性和阳性;未经DTT处理的抗体筛选红细胞、抗体鉴定细胞和献血者红细胞与患者血浆反应均呈阳性;DTT处理后的抗体筛选红细胞、抗体鉴定细胞和献血者红细胞与患者血浆反应均呈阴性;平行对照用的O型K(+)E(+)红细胞经DTT处理后转变为O型K(-)E(+)。结论抗-CD38单克隆抗体治疗多发性骨髓瘤,可干扰患者的血型血清学实验,用0.2 mol/L的DTT灭活红细胞表面的CD38抗原后,可以较好的去除这种干扰。     

抗-cE及抗-Jk^b和抗-Mur混合抗体致疑难配血分析——附1例报告

目的探讨1例疑难交叉配血不合的原因。方法通过ABO血型鉴定、Rh分型、直接抗人球蛋白试验、不规则抗体筛查、抗体特异性鉴定及抗体效价测定对患者配血不合进行分析。结果对照谱细胞反应格局,患者血清中检出抗-cE、抗-Jk^b和抗-Mur。结论患者血清中不规则抗体是导致交叉配血不合的主要原因。     

IgG-Jk~a联合IgG-cE、IgG-Fy~b抗体鉴定及输血策略分析

目的鉴定1例有复杂不规则抗体的标本,并探讨鉴定此类复杂不规则抗体的思路和方法及输血方案。方法对标本鉴定ABO、Rh、Duffy和Kidd等血型,采用盐水方法和间接抗球蛋白方法进行抗体筛选及鉴定抗体类型,采用盐水法、凝聚胺法和间接抗球蛋白方法进行交叉配血。结果患者血型血清学结果为O,CCDee,Jk(a±b+), Fy(a+b-);基因型结果为Jk(a-b+)和Fy(a+b-)。抗体鉴定结合交叉配血试验共检出血清中存在IgG-c、IgG-E、IgG-Fyb、IgG-Jk~a抗体,筛选以上4种抗原均为阴性的供血者与患者进行交叉配血相合后输注。结论对于复杂不规则抗体的鉴定,应当明确鉴定的思路,将多种实验技术和方法进行组合和分析,有效提高实验室解决疑难问题的能力,为临床提供相合血液,保障输血安全。     

NEC患儿T抗原暴露导致红细胞多凝集现象的鉴定及输血策略

目的 探讨坏死性小肠结肠炎（necrotising enterocolitis,NEC）患儿发生T抗原暴露导致红细胞多凝集现象的鉴定方法,为其制定输血策略,为指导临床为患儿科学、合理用血提供参考依据。方法 在本中心进行疑难交叉配血的2例NEC患儿,采用试管法对其进行ABO血型和Rh血型鉴定、直接抗人球蛋白试验（directantiglobulintest,DAT）、抗体筛查试验及交叉配血试验,采用3例ABO同型献血者血浆,3例AB型献血者血浆,3例同型脐血血浆等方法发现其出现红细胞多凝集现象,并用花生凝集素、MN血型抗体鉴定试剂及凝聚胺方法鉴定其多凝集现象。结果 两例患儿红细胞抗体筛查实验阴性;DAT实验阴性;交叉配血实验主侧阴性;次侧盐水法阳性,抗人球蛋白实验阳性;患儿红细胞与献血者同型血浆、AB型血浆呈阳性反应,与同型脐血血浆呈阴性反应;与花生凝集素均呈阳性反应患儿MN血型抗原均为阴性,患儿红细胞加入凝聚胺试剂不出现预期凝集现象。结论 NEC患儿出现多凝集红细胞现象表现为DAT实验阴性,但是次侧交叉配血不相合现象。此时应采用多种方法鉴定其红细胞发生多凝集现象,准确识别患儿红细胞多凝...     

微柱凝胶抗球蛋白法、凝聚胺法与固相凝集法检测在临床输血中的应用价值

目的 分析微柱凝胶抗球蛋白法（MGCT）、凝聚胺法及固相凝集法用于临床输血中的价值。方法 选取2022年9月至12月在东莞康华医院进行输血治疗的2 071例患者。其中血小板输注无效的106例患者通过固相凝集法测定血小板抗体,后对血小板抗体阳性的患者进行谱细胞检测并以谱细胞检测结果为金标准,分析固相凝集法和谱细胞检测在血小板抗体测定中的阳性符合率;所有患者通过MGCT及凝聚胺法开展配血,比较两种方法的交叉配血试验不合检出情况,并提出相应的解决方法。结果 血小板输注无效的106例患者通过谱细胞法测定血小板抗体显示：阳性30例,阴性76例;固相凝集法结果显示：阳性29例,阴性77例,阳性符合率为96.67%。2 017例受血者开展交叉配血试验显示64例存在交叉配血试验不合的情况,其中MGCT显示58例存在交叉配血试验不合的情况,符合率为90.63%;凝聚胺法显示50例存在交叉配血试验不合的情况,符合率为78.13%。结论 固相凝集法应用到临床输血期间血小板抗体检测中和谱细胞检测的符合率较高,MGCT用于交叉配血试验较凝聚胺法效果更为理想,能更好检出交叉配血试验不合的情况,值得应用。     

抗-JK~a抗体致疑难配血病例分析

河南大学附属郑州颐和医院收治抗-JK~a抗体引起不规则抗体阳性及疑难配血患者1例。常规用微柱凝胶卡式法对患者进行ABO、Rh(D)血型鉴定、直接抗人球蛋白试验、不规则抗体筛查及鉴定、吸收放散试验以及Kidd血型分型,最后用多种方法交叉配血,筛选出相合血液制品进行临床输注。患者血型为O型Rh(D)阳性,直接抗人球蛋白试验结果阴性,不规则抗体鉴定出特异性抗JK~a抗体,Kidd分型结果Jk(a-b+)。鉴定不规则抗体特异性可以为患者筛选合适的供血者,确保临床输血安全。     

1 例骨髓增生异常综合征多次受血患者血清存在抗-M，抗-cE 和抗-Jkb 抗体的鉴定及配血输血策略

目的 通过对临床送检的1 例骨髓增生异常综合征（myelodysplastic syndrome, MDS）多次受血患者的抗筛阳性标本进行鉴定,探讨此类复杂不规则抗体的鉴定思路、方法及输血方案。方法 鉴定标本ABO,Rh,MNS 和Kidd 等血型,应用2 种谱细胞及筛选出的无偿献血者细胞配合吸收放散实验进行抗体筛查、复杂不规则抗体特异性鉴定,采用盐水法、凝聚胺法和间接抗球蛋白方法进行交叉配血。结果 患者血型血清学结果为O,CCDee,M-N+ 和Jk(a+b-),其血清中检出IgM-M,IgG-cE 和IgG-Jkb 抗体,筛选M,c,E 及Jkb 抗原均为阴性的供血者与患者进行交叉配血,配血相合后进行输注。结论 针对多次受血患者产生的复杂不规则抗体,应灵活运用灵敏度高的血清学方法,联合多种谱细胞,准确鉴定出抗体特异性,为临床提供相合的血液,有效保障输血的安全性。     

异基因造血干细胞移植后并发自身免疫性溶血性贫血患者产生类抗体血清学检测方法及输血策略

目的探讨异基因造血干细胞移植（AHSCT）后免疫介导的自身免疫性溶血性贫血（AIHA）患者交叉配血不合的原因,血浆中不规则抗体筛选试验阳性时,不规则抗体鉴定的血清学检测方法的选择,处理措施及输血策略。 方法一例AHSCT后的3岁男性患儿因重度贫血为改善贫血症状需输注红细胞入住四川省人民医院。通过盐水试管法确定了患儿ABO、Rh血型后进行交叉配血试验发现患儿与多个同型供者血液不相合,考虑到患儿贫血严重,于是采用直接抗球蛋白试验来判断红细胞是否被致敏;通过盐水介质试管法、微柱抗球蛋白法和聚凝胺法进行ABO血型系统以外的不规则抗体筛选试验以确定血浆中有无不规则抗体;根据不规则抗体筛选试验结果选择聚凝胺法来确定抗体的特异性;通过盐水介质试管法、经典抗球蛋白法和聚凝胺法进行交叉配血实验,结合抗体鉴定的结果,综合分析选择合适的供者红细胞输注。 结果本例患儿ABO血型为AB型,Rh分型为CCDee,ABO血型已转变为供者血型。直接抗球蛋白试验强阳性,红细胞被抗体致敏。血浆中检出了不规则抗体,红细胞上放散下来的致敏抗体与血清中检出的不规则抗体均为类抗-Ce自身抗体。选择了避开类抗-Ce抗体的Ce抗原阴性的AB型红细胞输注后血色素升高,3 d后复查血常规Hb为79 g/L,输血有效。 结论任何类型的AHSCT后都有可能发展成AIHA,也就是血浆中可能存在某种自身抗体（类抗体）而破坏自身红细胞,若能够明确患者血清中存在的类抗体,即可避开这种抗体而筛选红细胞无相应抗原的供者,也就避免发生溶血性输血反应以安全输血。     

红细胞血型系统IgG型不规则抗体固相凝集法检测试剂盒的研发

目的研发基于固相凝集技术的红细胞血型系统IgG型不规则抗体检测试剂盒并评估其性能。方法采用单克隆抗人-RBC抗体包被96孔微板,按100μL/孔加入红细胞悬液形成细胞单层,经过裂解后形成红细胞膜抗原分子单层,加入保护液冷冻干燥后制备成检测用的反应微板;用梯度倍比稀释的多克隆羊抗球蛋白试剂(IgG+C3d/4)与IgG抗-D致敏的O型红细胞反应,筛选构建最佳指示系统;用不同批次的试剂盒检测低效价的IgG抗-D和抗-E,评估试剂盒在保存期内的抗原稳定性;与微柱凝胶抗球蛋白卡、进口同类试剂盒检测不同效价的抗体以测试3者的灵敏度;与进口同类试剂盒对350例血样标本进行检测,以明确2者的检测效能是否一致。结果 20μg/mL的抗人-RBC抗体溶液能够很好的固定红细胞膜抗原分子单层;稀释16倍的多克隆羊抗球蛋白试剂与致敏红细胞构建的指示系统指示效果最佳;试剂盒中冻干的红细胞膜抗原在6个月的保存期内保持稳定;本研究的试剂盒检测灵敏度高于Capture-R Ready Screen试剂盒和抗球蛋白卡;试剂盒与Capture-R Ready Screen对血样标本检测的阳性一致性百分率为98.0%,阴性一致性百分率为99.66%,总一致性百分率为99.43%。结论成功研发了基于固相凝集技术的用于红细胞IgG型同种免疫性不规则抗体检测的试剂盒,该试剂盒具有灵敏度好、保存期长、性能稳定等优势,与进口同类试剂盒等效。     

Capture-R法与微柱凝胶法在不规则抗体筛查中的应用

目的探讨Capture-R法与微柱凝胶法在不规则抗体筛查中的临床应用价值。方法采用微柱凝胶法与Capture-R法对600份临床患者标本做不规则抗体筛查平行试验,对抗体筛查阳性标本进行抗体鉴定。分析比较两种方法在不规则抗体筛查试验中的灵敏度和特异性。结果微柱凝胶法和Capture-R法阳性检出率分别为3.17%（19/600）和2.83%（17/600）,两种方法灵敏度比较差异无统计学意义（P〉0.05）;特异性分别为89.47%（17/19）和100.00%（17/17）,两种方法特异性比较差异无统计学意义（P〉0.05）。结论 Capture-R法在不规则抗体筛查试验中的灵敏度和特异性均达到微柱凝胶法检测水平,可有效检出有临床意义的红细胞抗体,提高临床输血安全。     

血型不规则抗体检测的方法学评价

目的寻找简便、快速、敏感的用于红细胞不规则抗体筛选的试验方法。方法收集6例临床疑难病例样本和1份IgG型试剂,采用盐水法、木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法平行检测患者血清中不规则抗体的效价,比较5种方法的敏感性、特异性。结果盐水法仅检出IgM类不规则抗体;木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法在检测IgG类抗体效价时差异不大,木瓜酶法的敏感性略低,微柱凝胶法对弱阳性IgG抗体检测的敏感性最高,但4种方法间差异无统计学意义（P〉0.05）,相关性较好（r〉0.99）。结论盐水法不能检出IgG抗体;木瓜酶法检测IgG抗体具有一定的局限性;凝聚胺法简便、快速,但为手工操作,不易标准化;抗人球蛋白法需要反复洗涤红细胞,费时,不便于开展大规模筛查;微柱凝胶法操作简便迅速、敏感性高、重复性好、易于标准化,适于临床输血前和孕妇产前大批量抗体筛选,是检测IgG抗体最为敏感的方法。     

1例输血患者疑难血型的鉴定与分析

目的鉴定与分析1例配血困难患者的ABO血型,为临床疑难血型的鉴定提供案例参考。方法采用血型血清学方法进行ABO、Rh血型、直接抗人球蛋白试验、不规则抗体筛查和抗体鉴定、吸收放散试验及抗体效价测定、唾液血型物质检测、交叉配血等试验,对1例血型鉴定疑难患者进行确诊。结果该例患者正定型为A型Rh阳性,反定型为AB型Rh阳性,进一步采用多种方法进行鉴定,最终确定为ABx亚型,经筛选后交叉配血相合。患者输注悬浮红细胞后,无不良反应。结论采用多种血清学方法对ABO血型正反不一致的标本进行血型鉴定,从而提高亚型检出率。     

Activation of human platelets by antibodies to thymocytes and beta 2-microglobulin. III. Effect of cell absorptions on the platelet aggregating and lymphocytotoxic activities of HATG and SA beta 2mG

Horse anti-human thymocyte globulin (HATG) and sheep anti-human beta-2-microglobulin globulin (SA beta 2-mG), both known to have immunosuppressive potency, were quantitatively absorbed by a number of human cells, then, tested for platelet aggregating and lymphocytotoxic reactivities. Peripheral blood lymphocytes, lymphoblastoid T- and B-cell lines, platelets and spermatozoa could remove varying amounts of both reactivities from HATG and SA beta 2mG. Daudi cells were effective in reducing the lymphocyte and some platelet reactivity of HATG but did not affect those of SA beta 2mG. Erythrocytes had no absorption capacity for either antibody. The qualitative and quantitative differences in absorbing with various cells on the platelet aggregating and lymphotoxic activities of HATG and SA beta 2mG indicate the high specificity and direct action of the antibodies on platelets and lymphocytes.     

Pre-transfusion testing problems caused by anti-lymphocyte globulin and their solution

Anti-lymphocyte globulin (ALG) is an antibody to human lymphocytes used to decrease T-cells in renal transplant patients. We recently encountered serologic problems in testing blood from patients treated with ALG. Thirty-nine patients undergoing acute kidney rejection developed positive direct and indirect antiglobulin tests following the administration of equine ALG. Sera from these patients reacted with all red cells (RBCs) tested using both polyspecific and monospecific anti-IgG anti-human sera. Eluates prepared from the patients' RBCs showed similar reactivity. The ALG panagglutinin did not react by manual hexadimethrine bromide (Polybrene) technique. The ALG panagglutinin could be neutralized by anti-human globulin. In our hands, these techniques were useful in distinguishing ALG panagglutinin from co-existing alloantibodies.     

Column agglutination technology: the antiglobulin test

A new system for typing and screening blood, based on the sieving effect of glass bead microparticles, has been developed. The test is performed in a microcolumn in which the red cell agglutinates are trapped in the glass bead matrix during centrifugation, and unagglutinated cells form a pellet at the bottom of the column. Anti- human globulin reagents were incorporated in the diluent and the new test system, column agglutination technology, was compared to conventional tube tests and low-ionic-strength method. Sera and plasmas (228 samples) were screened for red cell antibodies with two anti-human globulin reagents: one containing only anti-IgG and the other containing both anti-IgG and anti-C3b, -C3d. After initial testing, there was 94-percent agreement between column agglutination technology and tube tests, and after repeat testing, there was 97-percent agreement. The column agglutination technology anti-human globulin test eliminates the need to wash red cells, which decreases the overall test time. The test is easy to perform, and the results are more objective than those with tube and microplate methods.     

血型不规则抗体检测的方法学评价

目的寻找简便、快速、敏感的用于红细胞不规则抗体筛选的试验方法。方法收集6例临床疑难病例样本和1份IgG型试剂,采用盐水法、木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法平行检测患者血清中不规则抗体的效价,比较5种方法的敏感性、特异性。结果盐水法仅检出IgM类不规则抗体;木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法在检测IgG类抗体效价时差异不大,木瓜酶法的敏感性略低,微柱凝胶法对弱阳性IgG抗体检测的敏感性最高,但4种方法间差异无统计学意义(P>0.05),相关性较好(r>0.99)。结论盐水法不能检出IgG抗体;木瓜酶法检测IgG抗体具有一定的局限性;凝聚胺法简便、快速,但为手工操作,不易标准化;抗人球蛋白法需要反复洗涤红细胞,费时,不便于开展大规模筛查;微柱凝胶法操作简便迅速、敏感性高、重复性好、易于标准化,适于临床输血前和孕妇产前大批量抗体筛选,是检测IgG抗体最为敏感的方法。     

Comparative sensitivity of solid phase versus PEG enhancement assays for detection and identification of RBC antibodies.

Blood banks require a sensitive, specific, and efficient method to detect clinically significant RBC antibodies. Solid phase antibody screening methods are popular due to high sensitivity and automation. However, the high degree of reactivity detects "false positive" antibodies of questionable clinical significance leading to additional testing. We studied positive rates of Capture-R vs. PEG methods and categorized RBC antibodies identified by initial test results of 33,564 consecutive samples by Capture-R method. Capture-R was positive in 1,084/33,564 (3.2%) of samples. Using PEG as our "gold standard", PEG confirmed true positivity (i.e., > or = 1 cell reacting) in 710 Capture-R positive samples (65.5%); 374 Capture-R positive samples (34.5%) did not react in PEG (i.e., false positives). Of the 710 samples with true positivity, only 510 showed clinically significant alloantibodies. Using PEG as our "gold standard", only 2/3 of reactions by Capture-R were considered true positives. Because of ease and automation, Capture-R is popular as a screening test, but a more specific method may be helpful in order to identify truly significant alloantibodies.     

Serologic evidence that factor IX inhibitor in the plasma of hemophilia B patients detects factor IX on normal red cells

BACKGROUND: Patients with hemophilia B lack factor IX (F IX). These patients may become alloimmunized after the transfusion of F IX concentrates and may develop F IX inhibitors, which have been characterized as polyclonal IgG4 alloantibodies. Two cases in which F IX inhibitors caused difficulty in compatibility testing and antibody identification were encountered. It was hypothesized that, because F IX is present in normal plasma, it might be adsorbed by red cells in vivo and then be detected during antibody screening tests with serum containing F IX inhibitors. CASE REPORT: Sera from two African American half-brothers with hemophilia B were incompatible with all common and rare red cell phenotypes tested in the anti-human globulin test, but did not react with each other's red cells. The brothers' red cell antibodies were neutralized with both normal plasma and a commercially available F IX concentrate, which indicated that the red cell incompatibility was most probably caused by their F IX inhibitors. Red cells from an unrelated patient with hemophilia B and a very low titer of F IX inhibitor were tested against the half-brothers' sera and did not react. The compatible red cells from one of the half-brothers and the unrelated patient with hemophilia B adsorbed F IX from normal plasma or F IX concentrate after 37 degrees C incubation; this rendered them incompatible with the plasma containing F IX inhibitor from the other half-brother. CONCLUSION: F IX appears to be present on normal red cells and may be detected during compatibility and antibody identification procedures when serum or plasma containing F IX inhibitors is tested.