抗I抗体引起ABO血型正反定型不符及原因分析北大核心CSCD

目的探讨患者血清中抗I抗体的血清学特点及其对输血相容性检测的影响。方法在输血相容性检测中,出现正反定型不符和交叉配血不合的患者血标本,将其红细胞用37℃生理盐水反复洗涤后,进行复查血型和直接抗人球蛋白试验;对其血清进行冷凝集素吸收试验和不规则抗体筛选,对抗体筛选阳性的患者血标本进行抗体特异性鉴定、抗体性质及抗体效价测定。结果 6例患者ABO血型鉴定结果为:A型2例、B型3例、O型1例。患者血清标本在输血相容性检测中与反定型红细胞、自身红细胞、成人O型红细胞,在4℃、22℃发生1+～2+意外凝集,与脐血O型红细胞不发生凝集,与抗体筛选细胞在37℃不凝集,说明该抗体为抗I抗体。患者直接抗人球蛋白试验抗Ig G阴性、抗C3d阳性。将患者血清用2-巯基乙醇处理后,再与成人O型细胞、脐血O型细胞在4℃、22℃进行抗体检测,结果均不凝集,说明该抗体性质为Ig M型。抗体效价4℃为1∶128～1∶512、22℃为1∶32～1∶64、37℃为0。结论对输血相容性检测中出现意外凝集的患者血标本,采用37℃生理盐水洗涤红细胞、抗人球蛋白试验、冷凝集素吸收试验、不规则抗体筛选及特异性鉴定;并在37℃条件下进行血型鉴定和交叉配血试验,可保证检测结果的准确性和输血安全。     

抗I抗体引起ABO血型正反定型不符及原因分析

目的探讨患者血清中抗I抗体的血清学特点及其对输血相容性检测的影响。方法在输血相容性检测中,出现正反定型不符和交叉配血不合的患者血标本,将其红细胞用37℃生理盐水反复洗涤后,进行复查血型和直接抗人球蛋白试验;对其血清进行冷凝集素吸收试验和不规则抗体筛选,对抗体筛选阳性的患者血标本进行抗体特异性鉴定、抗体性质及抗体效价测定。结果 6例患者ABO血型鉴定结果为:A型2例、B型3例、O型1例。患者血清标本在输血相容性检测中与反定型红细胞、自身红细胞、成人O型红细胞,在4℃、22℃发生1+～2+意外凝集,与脐血O型红细胞不发生凝集,与抗体筛选细胞在37℃不凝集,说明该抗体为抗I抗体。患者直接抗人球蛋白试验抗Ig G阴性、抗C3d阳性。将患者血清用2-巯基乙醇处理后,再与成人O型细胞、脐血O型细胞在4℃、22℃进行抗体检测,结果均不凝集,说明该抗体性质为Ig M型。抗体效价4℃为1∶128～1∶512、22℃为1∶32～1∶64、37℃为0。结论对输血相容性检测中出现意外凝集的患者血标本,采用37℃生理盐水洗涤红细胞、抗人球蛋白试验、冷凝集素吸收试验、不规则抗体筛选及特异性鉴定;并在37℃条件下进行血型鉴定和交叉配血试验,可保证检测结果的准确性和输血安全。     

冷凝集素综合征的血清学特性及检测方法探讨

目的:了解冷凝集素综合征在不同疾病患者中的分布,冷凝集素的血清学特性及检测方法。方法:检测498例患者血清中的冷凝集素的效价和免疫球蛋白类型。对含有高效价冷凝集素的血清在4℃进行吸收,对经43℃的生理盐水洗涤后仍有凝集的RBC,采用45℃放散-聚蔗糖分离-微柱凝胶法进行血型鉴定和交叉配血试验。结果:在498例患者血清中,含有冷凝集素者324例(65%),其中冷凝集素的效价≤1∶32者435例(87%),≥1∶64者60例(12%)。冷凝集素综合征在血液病或重度贫血患者中占17.6%,在肿瘤患者中占13%,在其他疾病患者中占6%。冷凝集素的免疫球蛋白类型均为IgM。用45℃放散-聚蔗糖分离-微柱凝胶法检测冷凝集素效价增高者的ABO血型,正反向定型的结果一致,交叉配血试验无假凝集现象。结论:患者中冷凝集素综合征占12%,冷凝集素的效价增高时可干扰ABO血型鉴定和交叉配血试验的结果,采用4℃吸收冷凝集素,用43℃的生理盐水洗涤RBC,以45℃放散-聚蔗糖分离-微柱凝胶法,能正确鉴定ABO血型,保证交叉配血试验结果的准确性。     

微柱凝胶法检测疾病对ABO血型抗原的影响及血清学特性分析

目的：探讨疾病对ABO血型抗原性的影响及其血清学特性分析。方法：应用微柱凝胶法、单克隆抗体血型分型试剂和RBC试剂，对100例患者和30例献血员的RBC（检测抗原）和血清（检测抗体）进行正反定型，测定血型抗原性和抗体效价。对血型抗原性极度减弱导致血型鉴定困难的患者血液，进行吸收放散、血型物质测定、不规则抗体筛选和抗球蛋白试验。结果：100例患者中血型抗原性减弱者占24％，其中自血病患者占40％（12／30），恶性肿瘤患者占36．67％（11／30），其他疾病患者占2．50％（1／40）。抗原性减弱在ABO血型中的分布：A型为40．48（17／42），B型为16．67％（5／30），O型为7．14％（2／28）；献血员血型抗原性减弱者为3．33％（1／30）。100例患者中血清血型抗体效价降低者2％，献血员血清血型抗体效价降低者为0％。结论：白血病和恶性肿瘤对血型抗原性的影响较其他疾病大，患者ABO血型抗原性减弱正定型时容易导致血型误判，但其血清中仍存在着规则的抗A或抗B抗体，唾液中含有血型物质，应用微柱凝胶法进行正反定型，同时进行吸收放散试验和血型物质的测定，能提高血型检测的准确性.     

血型抗体减弱或缺乏导致ABO血型正反定型不相符的检测分析

目的分析血型抗体减弱或缺乏的原因及其对ABO血型鉴定的影响和处理方法。方法采用正反定型、抗体增强方法、吸收放散试验、抗球蛋白试验、不规则抗体筛选及交叉配血试验对患者进行血型血清学检测。结果病例1、 2、 3、 4正定型为O型,病例1、 2反定型抗A、抗B抗体在常规方法中与Ac、 Bc不凝集,病例3抗B抗体在常规方法中与Bc不凝集。经采用抗体增强方法及吸收放散试验,使抗A、抗B抗体凝集强度增加到弱凝集至2+。病例4正定型为O型、病例5正定型为A型,反定型采用抗体增强方法及吸收放散试验,仍未检测出抗B抗体,为抗B抗体缺乏。结论患者血型抗体减弱或缺乏可导致ABO血型正反定型不相符,使判定结果和临床及时输血受到影响。     

抗Mur抗体的血清学检测及其临床意义

目的 分析抗Mur抗体的血型血清学检测结果及其临床意义。方法 采用微柱凝胶法(MGT)对2例患者血标本进行ABO、 RhD血型鉴定及不规则抗体筛选,结果有疑问时再采用试管盐水法(NS)、直接抗人球蛋白试验(DAT)、间接抗人球蛋白试验(IAT)、吸收放散试验、抗体特异性鉴定及交叉配血试验进行检测鉴定。结果 患者1 ABO血型为A型,RhD阳性,DAT阴性。患者2ABO血型为B型,RhD阳性,DAT阴性。两例患者在采用NS、 MGT进行血型血清学检测中,交叉配血不合。经血型血清学检测鉴定为IgM+IgG型抗Mur抗体,抗体效价为1∶1～1∶4,凝集强度为2+～1+。结论 抗Mur抗体大多数为IgM型抗体,极少数为IgG型抗体,或IgM型+IgG型混合抗体,该抗体可引起正反定型不符或交叉配血不合,在37℃及IAT有反应时,可引起溶血性输血反应和新生儿溶血病的发生。     

产前血型IgG抗体水平的检测及其临床意义

目的探讨ABO血型不合孕妇的产前IgG抗体水平,了解IgG抗体效价异常在孕妇中所占比率及临床意义,为预防及诊治新生儿溶血病（HDN）采取有效的防治措施。方法用抗人球蛋白试管凝集法进行IgG抗A或抗B的ABO血型抗体效价检测。结果910例孕妇中,血清效价大于64者有108例,异常检出率为11.9%。IgG抗A效价大于64者有64例,检测率为11.8%;检测IgG抗B效价大于64者有54例,检测率为14.1%。讨论妊娠中IgG抗体效价与新生儿溶血密切相关,ABO血型不合的孕妇应及时作产前血清学的检测,可预防新生儿溶血病的发生及减轻胎儿受害的程度。     

B亚型患者血清不规则抗B抗体的检测分析

目的 分析B亚型(Bw)患者血清中存在IgM型不规则抗B抗体的血型血清学检测结果及其临床意义。方法 对ABO血型正反定型不一致或交叉配血不相合的患者血标本,采用抗A、抗B、抗AB、抗H定型试剂和不同试验方法进行正反定型,采用吸收放散试验、直接抗人球蛋白试验及ABO血型以外不规则抗体筛查进行检测鉴定。对需要输血的B亚型及血清中存在不规则抗B抗体的患者,选择O型悬浮红细胞或洗涤红细胞进行配合性输血治疗。结果 15例患者红细胞与抗B定型试剂在22℃立即离心,凝集强度弱于1+(1+^w),置4℃反应15 min,离心2次,凝集强度强于1+(1+^s)～2+(2+^s)。患者红细胞与抗AB定型试剂的凝集强度大于抗B定型试剂,与抗H定型试剂的凝集强度小于O型红细胞。在试管法反定型中,患者血清与B型试剂红细胞混合,在22℃立即离心,凝集强度为1+^w～1+^s;置4℃反应15 min,离心2次,凝集强度为1+^s～2+。结果显示患者血清中存在凝集强度为1+～2+的IgM型不规则抗B...     

76例Lewis血型抗体血清学特征及其对输血相容性检测的影响

目的探讨Lewis血型系统抗体的血清学特征及其对输血相容性检测结果的影响和处理方法。方法采用微柱凝胶法对临床送检的患者血标本进行ABO及Rh血型鉴定,采用Liss/Coombs卡进行不规则抗体筛选及交叉配血试验,对抗体筛选阳性及交叉配血不合的血标本采用微柱凝胶法和盐水试管法进行抗体特异性鉴定,对有特异性抗体者采用盐水试管法进行Lewis血型抗原检测。结果在76例Lewis血型抗体中,抗Lea抗体75例(98.7%),抗Leb抗体1例(1.3%);男26例(34.2%),女50例(65.8%),女性患者抗体阳性率明显高于男性。76例Lewis血型抗体在盐水试管法中出现凝集76例(100%),在Liss/Coombs卡中出现凝集72例(94.7%),Lewis血型抗原检测均为Lea-b-。76例Lewis血型抗体在抗体筛选及交叉配血中均出现凝集,13例在ABO血型反定型中出现凝集。盐水试管法凝集强度为1+以下～1+(22℃),Liss/Coombs卡凝集强度为混合外观凝集至1+以下(37℃),抗体类型为IgM型。结论产生Lewis血型抗体的血型抗原均为Lea-b-;Lewis血型抗体可干扰输血相容性检测结果,采用盐水试管法和Liss/Coombs卡联合检测可提高对Lewis血型抗体的检出率。     

H抗原缺失患者抗H抗体血型血清学检测及结果分析

目的分析H抗原缺失患者抗H抗体血型血清学检测结果及其对输血相容性检测的影响。方法采用血型血清学方法对出现正反定型不符及交叉配血不合的3例患者血标本,进行抗体筛选及特异性鉴定,唾液血型物质测定,对患者红细胞进行H抗原检测及直接抗人球蛋白试验。结果患者中A型1例, AB型2例, RhD为阳性。输血相容性检测在4℃、 22℃及37℃出现4+～±的凝集反应,引起ABO血型正反定型不符或交叉配血不合,患者红细胞H抗原及直接抗人球蛋白试验阴性。经血型血清学鉴定为抗H抗体,抗体类型为IgM型,抗体效价4℃为1∶64～1∶128, 22℃为1∶4～1∶32, 37℃为1∶1～1∶2。抗体效价及凝集强度随温度上升而降低。结论 22℃有反应活性的抗H抗体,可干扰输血相容性检测结果, 37℃有反应活性的抗H抗体可能引起输血反应。含有抗H抗体的患者应输同型血,如紧急抢救无同型血而需输O型红细胞时,应用输注盐水法和抗人球蛋白法交叉配血均相合的血液。     

Retrospective analysis of molecular biology mechanism of ABO blood group typing discrepancy among blood donors in Jinan blood station

BackgroundTo accurately identify ABO blood typing in pre-transfusion testing is very important to ensure blood transfusion safely, which is a major responsibility of blood station.MethodsEighty-one blood donors samples with ABO blood group typing discrepancy was collected among 61952 donor samples in our blood station from January 2019 to July 2020. Blood group serological method was used to detect ABO blood group. DNA Sequencing was used to determine the genotype. The antibody screening test detects antibodies other than ABO.ResultsIn total, 61,952 donor samples were analysed for ABO typing discrepancies. The incidence among blood donors was 0.13% (81/61952). The most common reason of ABO typing discrepancies was due to specific antibody or non-specific agglutination (54.32%, 44/81), mainly anti-M antibody, cold autoantibody, anti-D antibody, anti-N antibody and anti-Lea antibody. The major cause of forward typing discrepancies among blood donors was ABO subgroups (25.93%, 21/81), including 10 cases of A subtype (1 case of A2, 2 cases of A3, 2 cases of Ax, 3 cases of AxB, 1 case of Ael, 1 case of Ahm), 6 cases of B subtype (2 cases of B3, 1 case of Bel, 3 cases of AB3), 2 cases of B subtype (A), 1 case of cisAB, and 2 cases of acquired B. The serum antibody was weakened in 16 cases (19.75%).ConclusionsThe blood types should be correctly identified by combining serology with gene sequencing to ensure the safety of clinical blood transfusion, when the forward and reverse typing discrepancies among the blood donors.     

用孕妇外周血游离DNA检测胎儿ABO血型

目的探讨用孕妇血浆中游离DNA检测胎儿ABO血型方法的可行性。方法应用DNA提取试剂盒(qiagen德国)从46例9～22周孕妇血浆提取胎儿DNA,采用复合PCR-RFLP检测胎儿ABO血型,并用常规血清学方法检测母体及新生儿血型。结果 46例样本中,产前检查能正确显示胎儿血型的有38例,总检出率为(38/46)82.61%。其中:①孕妇血型为O型,丈夫血型为A型16例,产前检测孕妇胎儿血型为A型,待婴儿出生后血清学检测出12例,符合率为(12/16)75.00%;②孕妇血型为O型,丈夫血型为B型占13例。产前检测孕妇胎儿血型为B型,待婴儿出生后血清学检测出9例,符合率为(9/13)69.23%。③孕妇血型为O型,丈夫血型为O型占11例,产前检测孕妇胎儿血型为O型,待婴儿出生后血清学检测出10例,符合率为90.90%。总检出符合率(10/11)81.58%。结论用孕妇血浆中游离DNA检测胎儿ABO血型方法可行,对诊断和预防新生儿ABO血型不合性溶血病具有积极意义。     

广州地区无偿献血者不规则抗体筛查结果分析

目的了解广州地区无偿献血者不规则抗体的频率、类型、特异性和抗体效价。为输血前检查策略的制定提供依据。方法随机抽取2012年11月至2013年3月广州血液中心无偿献血者血液样本20160例。选择含特定抗原的筛选红细胞,采用聚凝胺介质微板法进行不规则抗体初筛,阳性样本使用试剂筛选细胞和试管法进行确证试验,仍然阳性的样本采用谱细胞微柱法进行特异性鉴定并测定效价。结果20160名广州地区无偿献血者中共筛查出不规则抗体97例,检出率为0．48％,其中IgG型抗-E1例,IgM型抗体96例,包括抗-P14例,抗-M2例,抗-Lewisnn1例,上述抗体效价均不超过8;冷自身抗体59例和非特异性不规则抗体30例。女性不规则抗体阳性率显著高于男性（xz=18．7201,P=1．51E．05）。结论广州献血人群中存在低比例的不规则抗体,对献血者进行不规则抗体筛查有利于电子配血及血液预警系统的建立,对提升临床用血安全性、有效性和智能化水平有着重要意义。     

孕妇ABO血型抗体效价检测与治疗

目的探讨AB0血型抗体效价异常与不良孕产史的关系及治疗效果。方法采用盐水试管凝集法,进行IgG抗A或抗B的ABO血型抗体效价检测;对抗体效价异常者进行中西医结合治疗。结果 228例孕妇中,血清效价≥1:128者有183例,异常检出率为80.26%;通过治疗三个疗程治愈175例,治愈率95.63%;三个疗程显效5例,占2.73%;总有效率为98.36%,效果满意。结论对有自然流产、死胎、早产、死产及新生儿溶血等不良孕产史的、女方为O型血的夫妇,开展ABO血型抗体效价检测对预防和治疗母婴血型不合溶血病有积极作用。     

全自动血型仪检测ABO血型正反定型不符原因分析及处理策略

目的 探讨在鉴定ABO血型中全自动血型仪正反定型不符的原因,并探寻解决对策。方法 对2017年10月~2018年8月遂宁市中心医院血型仪检测的298例ABO血型正反定不符的标本进行分析,同时结合试管法、抗人球蛋白实验、吸收放散实验、不规则抗体鉴定结果及病史资料分析定型不符的原因并进行归类。结果 298例正反定型不符标本准确鉴定后归纳分为4类:可被纠正类型（标本、人员操作、试剂和仪器原因）占24.16%;抗原原因占7.38 %,其中包括抗原减弱以及ABO亚型;抗体减弱占31.88%,包括抗体合成不足（如婴幼儿）,抗体缺失（如老年人和肿瘤患者）,意外抗体等;其他原因占36.58%,主要包括自身免疫性溶血性贫血、冷凝集素综合症、高球蛋白血症等。结论 鉴定血型前标本质量控制、仪器定期的保养维护以及试剂红细胞的更换可以有效减少正反定型不符,抗体合成不足、自身免疫性疾病以及冷凝集素综合症是导致ABO正反定型不符的主要原因。建议院方根据可能原因进行相关确认实验,准确鉴定ABO血型从而保证患者输血安全。     

温州市区育龄妇女孕前巨细胞病毒感染现状调查

目的了解温州地区育龄妇女孕前人巨细胞病毒（HCMV）感染的状况。方法收集2008年10月至2010年6日参加温州市龙湾区免费孕前优生筛查的妇女血标本2869份,采用酶联免疫吸附试验（ELISA）检测血清HCMV IgG/IgM抗体;HCMV IgM抗体阳性标本,采用实时荧光定量聚合酶链反应（FQ-PCR）检测血HCMV DNA载量;HCMV IgG/IgM抗体双阳性标本,采用尿素变性结合ELISA技术检测IgG抗体亲和力指数（AI）。结果 2869份孕前妇女血清中HC-MV IgG抗体阳性检出率为97.77%（2805/2869）,HCMV IgM抗体阳性检出率为0.77%（22/2 869）,IgG/IgM抗体均阳性检出率占0.17%（5/2 869）;22份HCMV IgM阳性标本中,血HCMV DNA阳性检出率为68.18%（15/22）;5份HCMVIgG/IgM双阳性标本中,检出低亲和力IgG抗体1份,中等亲和力IgG抗体2份,高亲和力IgG抗体2份。结论温州市区育龄妇女孕前HCMV IgG抗体阳性率高;对HCMV IgM抗体阳性孕前妇女应进行多指标检测以判断HCMV感染的状态,为减少出生缺陷、做好优生优育服务提供依据。     

Natural erythrocyte agglutinins in the serum of the Australian freshwater catfish, Tandanus tandanus (Mitchell): I. Examination of the specificities of the agglutinins with emphasis on the ABH agglutinins

Normal serum samples from the Australian freshwater catfish, Tandanus tandanus (Mitchell), were shown to have haemolytic and haemagglutinating activity for a variety of animal erythrocytes. Specific agglutinins were present for the erythrocytes of most animals tested but some cross-reactivity of agglutinins was demonstrated.

Evidence was obtained that the greater part of the human erythrocyte agglutinating activity was directed toward antigens of the ABH system. Whereas some sera clearly showed anti-H or anti-A and B agglutination patterns, others agglutinated equally erythrocytes of different groups. ABO erythrocytes were agglutinated to higher titres than were erythrocytes of the `Bombay' phenotype, salivas from secretors inhibited the agglutination of erythrocytes carrying appropriate ABH antigens, whereas salivas from non-secretors did not and absorptions of sera often revealed ABH specific agglutinins.

While some sera agglutinated equally ABO erythrocytes because they contained a mixture of agglutinins with different ABH specificities, other serum samples containing one predominating species of agglutinin caused equal agglutination of O, A, AB and B erythrocytes. The specificity of this latter type of agglutinin is discussed.

     

A dispermic chimera with mixed field blood group B and mosaic 46,XY/47,XYY karyotype

Chimerism in humans is a rare phenomenon often initially identified in the resolution of an ABO blood type discrepancy. We report a dispermic chimera who presented with mixed field in his B antigen typing that might have been mistaken for the B3 subtype. The propositus is a healthy Korean male blood donor. Neither his clinical history nor initial molecular investigation of his ABO gene explained his mixed field agglutination with murine anti-B. Chimerism was suspected, and 9 short tandem repeat (STR) loci were analyzed on DNA extracted from blood, buccal swabs, and hair from this donor and on DNA isolated from peripheral blood lymphocytes from his parents. The propositus' red blood cells demonstrated mixed field agglutination with anti-B. Exon 6 and 7 and flanking intronic regions of his ABO gene were sequenced and revealed an O01/O02 genotype. B allele haplotype-specific PCR, along with exon 6 and 7 cloning and sequencing demonstrated a third ABO allele, B101. Four STR loci demonstrated a pattern consistent with a double paternal chromosome contribution in the propositus, thus confirming chimerism. His karyotype revealed a mosaic pattern: 32/50 metaphases were 46,XY and 18/50 metaphases demonstrated 47,XYY.     

Evaluation of a protein microarray method for immuno-typing erythrocytes in whole blood

All donor blood samples must be tested pre-transfusion to determine the blood type of donor erythrocytes, based on the ABO typing system. Current methods of testing are well characterised, but require a number of processing steps prior to analysis. In addition, standard testing protocols require additional assays such as hepatitis C and HIV testing be performed separately. We describe and evaluate a protein microarray platform for ABO blood typing that has the potential to be a simple reliable high throughput method, with the added capability for the integration of other important pre-transfusion tests. Sixty seven donor blood samples were incubated on microarrays printed with multiple spotted replicates of blood type antigen specific antibodies. We utilised a hold-out cross validation approach, combined with Receiver Operator Characteristic (ROC) curves to define thresholds within which a sample could be defined as being of a particular blood type. The threshold values from the ROC curve analysis demonstrated an excellent ability to accurately separate samples based on ABO blood type. The results obtained when the thresholds from the training sets were applied to test sets were also very encouraging, with misclassified samples being present in only 2 of the training sets and a mean classification error of 4.28%. When the mean thresholds were applied to the 67 donor samples, 95.5% were correctly blood typed (64 of 67 samples). We have demonstrated the ability of our protein microarray platform to successfully and accurately type human whole blood samples. We believe that this flexible platform provides a strong basis for an integrated approach for combined blood typing and pathogen testing in human whole blood.