Genomic analyses of KEL alleles in alloimmunized thalassemia patients from Iran

ObjectivesSerological methods are unreliable for red blood cells (RBCs) antigen typing in multi-transfused thalassemia patients due to the presence of donor RBCs in the recipient’s circulation and interfering antibodies. Kell blood group system is important in transfusion medicine and Kell antibodies have shown as the most prevalent antibodies in thalassemia patients. We intended to determine the genotype of Kell antigens among Iranian alloimmunized thalassemia patients using molecular methods and compare the results with serological phenotyping.MethodsTwo hundred thalassemia patients participated in this study. Blood group phenotype was performed by the serological method, while the genotype was determined for KEL*01, KEL*02, KEL*03, and KEL*04 alleles using PCR-Sequence Specific Primer (PCR-SSP) method. The genotypes of patients with incompatibility between phenotype and genotype were re-evaluated by Restriction Fragment Length Polymorphism-PCR (RFLP-PCR) and confirmed by DNA sequencing in all cases.ResultsTen patients were found with discrepancies between genotype and phenotype; however, there was a complete agreement between the results of SSP-PCR, RFLP-PCR, and DNA sequencing. Six discrepancies were found in the KEL*01/KEL*02 allele when serologically phenotyped as K-k+. One patient with K-k- and three patients with Kpa-Kpb + phenotype were identified as KEL*01/KEL*02 and KEL*03/KEL*04, respectively.ConclusionIt can be concluded that molecular genotyping is more reliable compared with the serological method, especially in the patients who have received multiple transfusions. Therefore, using a combination of these techniques can lead to a better matched transfusion in these patients.     

1例新的弱D型的鉴定

目的分析1例新的Rh血型弱D型个体的RHD等位基因及其红细胞D抗原表位。方法采用常规血清学方法检测Rh血型D、C、c、E和e抗原表型,间接抗人球蛋白试验(IAT)确认D抗原,并分析D抗原表位;序列特异性引物-聚合酶链反应(PCR-SSP)测定RHD基因,然后分析RHD编码区全长序列,并检测RHD杂合型。结果血清学显示该个例为D抗原弱表现型,Rh因子为D+C+c+E-e+,PCR-SSP检测RHD基因显示与正常Rh(D)阳性对照相同。RHD编码区序列分析发现第9外显子存在1 212C>A碱基突变,其余外显子序列则与正常RHD基因一致(GenBank EF103573),RHD合子型鉴定为RHD+/RHD-杂合型,提示该个体基因型为CDe/cde。红细胞D抗原表位分析显示其具有基本完整D抗原表位。结论该个例为RHD1 212C>A碱基突变形成弱D72型。     

Benefits of blood group genotyping in multi‐transfused patients from the south of Brazil

We evaluated the usefulness of blood group genotyping as a supplement to hemagglutination to determine the red blood cell (RBC) antigen profile of polytransfused patients with hematological diseases and renal failure. Seventy‐nine patients were selected. They all received more than three units of blood and eight (10%) had already clinical significant alloantibodies occurring alone or in combination against Rh, K, Fya, and Di antigens. DNA was prepared from blood samples and RHCE*E/e, KEL*01/KEL*02, FY*01/FY*02 and JK*01/JK*02 alleles were determined by using PCR‐RFLP. RHD*/RHD*Ψ and RHCE*C/c were tested using multiplex PCR. Discrepancies for Rh, Kell, Duffy, and Kidd systems were found between the phenotype and genotype‐derived phenotype in 16 of the 38 chronically transfused patients. The genotypes of these patients were confirmed by DNA array analysis (HEA Beadchip^?; Bioarray Solutions, Warren, NJ). Genotyping was very important for the determination of the true blood groups of the polytransfused patients, helped in the identification of suspected alloantibodies and in the selection of antigen‐negative RBCs for transfusion. J. Clin. Lab. Anal. 24:311–316, 2010. © 2010 Wiley‐Liss, Inc.     

SI35Blood Group Antigen Typing Using DNA Microarray BeadChipsTM–First UK Report

Background The determination of human blood group phenotypes has routinely involved red cell agglutination techniques. The genes for all but one of the 29 human blood group systems have now been cloned and the molecular bases of all clinically significant blood group polymorphisms are known. Consequently these DNA sequences can be used to predict blood group phenotypes. Molecular typing as an adjunct to serology offers improved accuracy and can save both rare reagents and laboratory time while screening for compatible donor units. This study determined the feasibility of analysis of single nucleotide polymorphisms (SNPs) associated with a variety of blood group antigens using a DNA microarray system with a beadchip technology platform. Method SNPs were tested by elongation‐mediated analysis of polymorphisms (eMAPTM) using allele‐specific oligonucleotides with variable 3’‐terminal sequences attached to colour‐encoded beads. The beads are assembled into arrays on semiconductor chips (BeadChip^TM), available from Bioarray Solutions. Elongation products for SNPs or small deletions/insertions were simultaneously detected by instant imaging of fluorescence signals from the entire array. Seventeen polymorphisms associated with Human Erythrocyte Antigens and one mutation associated with hemoglobinopathies (CO, DI, DO, FY, JK, KEL, LU, LW, MNS, SC and HgbS) were included in the HEA BeadChip^TM Kit. Results The system was validated using a reference DNA panel of known phenotype (n = 30) and genotype, established by conventional DNA‐PCR SSP assays. Replicate testing (n = 6 reps for eight samples) gave 100% consistent data demonstrating the Bioarray system to be both reliable and reproducible. The initial concordance rate with serology was 99.2%. Conclusion The BeadChip System represents the first integrated flexible array solution for high complexity transfusion testing, accommodating ‘multiplex’ analysis of polymorphisms. This high throughput, microarray system offers the potential to determine extended genotypes of patients and maintain an extended‐genotype, national panel of donors to provide compatible blood for sensitised patients. The initial high concordance rate and the rarity of CE marked phenotyping reagents justifies its further development.     

Clinical aspects of Rh genotyping

The Rh antigens are encoded by the RHD and RHCE genes. In RhD negative individuals the RHD gene is absent or grossly deleted. Routinely, Rh typing is performed by haemagglutination. However, there are some clinical situations in which serological techniques are not suitable for determining the red blood cell phenotype accurately. Most anti-D sera may not agglutinate erythrocytes possessing a reduced expression of the D antigen. In these cases, DNA-based analyses may be better than serological typing to infer the appropriate phenotype. Agglutination methods are also of limited use for determining the red blood cell phenotype of a foetus at risk of haemolytic disease of the newborn. Molecular RHD typing using amniocytes or DNA obtained from maternal plasma may obviate the need of amniocenteses during pregnancy when the foetus is RhD negative, thus providing an important tool in managing possible sensitization by foetal erythrocytes. Classical haemagglutination has limitation in patients with autoimmune haemolytic anaemia. Erythrocytes coated with IgG cannot be accurately typed for red blood cell antigens, particularly when directly agglutinating antibodies are not available or IgG removal by chemical treatment is insufficient. Molecular genotyping is very important for determination of the true blood group antigens of these patients. RHD genotyping with a specificity and sensitivity comparable to serologic methods is of practical importance to overcome the limitations of serology and, in addition, to improve the currently possible resolution.     

Red Blood Cell Antigen Genotyping for Sickle Cell Disease,Thalassemia, and Other Transfusion Complications

Since the discovery of the ABO blood group in the early 20th century, more than 300 blood group antigens have been categorized among 35 blood group systems. The molecular basis for most blood group antigens has been determined and demonstrates tremendous genetic diversity, particularly in the ABO and Rh systems. Several blood group genotyping assays have been developed, and 1 platform has been approved by the Food and Drug Administration as a “test of record,” such that no phenotype confirmation with antisera is required. DNA-based red blood cell (RBC) phenotyping can overcome certain limitations of hemagglutination assays and is beneficial in many transfusion settings. Genotyping can be used to determine RBC antigen phenotypes in patients recently transfused or with interfering allo- or autoantibodies, to resolve discrepant serologic typing, and/or when typing antisera are not readily available. Molecular RBC antigen typing can facilitate complex antibody evaluations and guide RBC selection for patients with sickle cell disease (SCD), thalassemia, and autoimmune hemolytic anemia. High-resolution RH genotyping can identify variant RHD and RHCE in patients with SCD, which have been associated with alloimmunization. In the future, broader access to cost-efficient, high-resolution RBC genotyping technology for both patient and donor populations may be transformative for the field of transfusion medicine.     

Discrepancies between red cell phenotyping and genotyping in daily immunohematology laboratory practice

False-positive and false-negative reactions exist for serological and molecular antigen typing methods. If the predicted phenotype is inconsistent with the patient`s known antibodies or serological phenotype, the discrepancy must be investigated. False-negative and false-positive results are clinically problematic in blood donors and patients. In this study, we investigated discrepant results between serology and molecular testing in patients and blood donors that occurred in daily molecular laboratory practice over a two year-period. SCD patients represented a large percentage of our cases of discrepancies but we also observed a high prevalence of discrepancies between phenotypes and genotypes in blood donors. The main reasons that led to discrepancies were recent transfusions and limitations of phenotyping. Discrepancies classified as false positive phenotype/true negative genotype and false negative phenotype/true positive genotype occurred mainly in patients with recent transfusions and individuals with RH variants while those classified as true negative phenotype/false positive genotype involved null phenotypes due to silent genes. Despite the limitations of molecular methods currently employed, we found more false-negative and false-positive phenotypes than genotypes demonstrating that genotyping is more efficient to define the blood types, especially in transfusion dependent patients.     

HLA-A33表型、TNF-α/TNF-RⅡ基因多态性和肠道病毒71型脑炎的相关性研究

[目的]研究HLA-A33表型,TNF-α/TNFRⅡ单核苷酸多态性与肠道病毒71型(EV71)感染遗传易感性的关系,探讨不同基因表型对EV71感染患病风险的影响.[方法]收集2009年9月至2010年10月EV71检测为阳性的急性期中枢神经系统感染患儿123例,根据病情轻重分成轻症组、重症组;提取患儿外周血白细胞的基因组DNA,用序列特异性引物-聚合酶链反应(PCR-SSP)技术检测EV71感染患儿及正常对照儿童HLA-A33表型、TNF-α-308位点的多态性和限制性片段长度多态性-聚合酶链反应(PCR-RFLP)检测TNFRⅡ+196位点的多态性.[结果]EV71感染患儿中HLA-A33表型阳性率(24.39％)与健康儿童组阳性率(11.82％)的差异有统计学意义(x2=6.099,P＜0.05).EV71中枢感染患儿TNF-α-308位点A等位基因频率明显高于正常对照组(x2=6.367,P＜0.05),感染组TNF-α-308GG基因型分布频率显著低于正常对照儿童(x2=5.393,P＜0.05);而轻症组与重症组相比,其差异无统计学意义(P＞0.05).TNFRⅡ+196位点T等位基因频率在EV71感染组与对照组、轻症组与重症组之间的差异均无统计学意义(P＞0.05).HLA-A33(+)患儿中,EV71感染组TNFRⅡ+196TG基因型频率明显高于正常对照组(x2=3.866,P＜005),而在HLA-A33(-)儿童中,其差异无统计学意义.[结论]HLA-A33表型、TNF-α-308位点基因多态性与EV71中枢感染有关,TNF-α-308GG基因型可能为儿童不易感染EV71的保护基因.TNFRⅡ+196位点基因多态性与EV71感染无关;TNFRⅡ+196TG基因型在一定程度上升高了HLA-A33(+)患儿EV71中枢感染的发病概率.     

MN基因定型在恶性血液病异基因造血干细胞移植植活证据检测中的临床研究

目的探讨MN基因型定型在恶性血液病异基因造血干细胞移植植活证据检测中简单可行的方法。回顾性分析了10例患者恶性血液病异基因造血干细胞移植的临床资料。方法分别采取受者和供者移植前及移植后14天及365天的血液标本,根据已知基因序列设计合成引物,采取PCR-SSP的方法检测方法的检测受者和供者移植后的基因型和基因亚型。结果 10例患者MN为5例,MM为3例、NN为2例或MGMG2例,MTMT1例,MGMT6例,MTN1例。与血清学结果完全吻合。显示两组样本的表型与基因型吻合很好,资料的说明性较好（P〉0.05）。结论血型基因检测的证据表明在同种异体造血干细胞移植中血型基因型可以作为早期监测的指标、标本用量少、科学敏感性强。     

Pattern of distribution of 35 red cell antigens in regular voluntary blood donors of South Gujarat,India

Background

Extended phenotyping is one of the important method of reducing red cell alloimmunisation. Extended phenotyping of red cells from voluntary donors have many uses in addition to its application in population genetics. As there was very little data extended phenotyping on a cohort of Indian Voluntary blood donors this project was undertaken.

Detection of blood group genes using multiplex SNaPshot method

BACKGROUND: The determination of blood group antigens in patients and donors is of primary importance in transfusion medicine. Blood group antigens are inherited and are polymorphic in nature. The majority of polymorphic blood group antigens arise from single-nucleotide polymorphisms (SNPs) in the blood group genes. Many DNA-based assays, such as species-specific polymerase chain reaction (PCR), PCR–restriction fragment length polymorphism, and microchips, have been described to study variant blood group genes. In this study, the SNaPshot (Applied Biosystems) method was adapted to detect SNPs in 10 common blood group systems.
STUDY DESIGN AND METHODS: DNA regions of interest were amplified in multiplex PCR and annealed to specific oligonucleotide probe primers of different lengths. AmpliTaq DNA polymerase extended the primers by adding only a single fluorescent ddNTP to its 3' end and was detected by differential mobility in capillary electrophoresis in a genetic analyzer. Results were analyzed using computer software in SNaPshot default analysis method.
RESULTS: Seventeen SNP sites in 29 blood samples, previously phenotyped and/or genotyped, were used to test the accuracy and reproducibility of multiplex SNaPshot assays. The results were compared with the previously analyzed types. SNaPshot analyses predicted the 17 SNP sites accurately for all the 29 blood samples. Both homozygous and heterozygous blood groups were detected with equal confidence.
CONCLUSION: Blood group detection by SNaPshot method is a practical alternative to antibody-dependent phenotype prediction. Starting with DNA, this method is fast with a turnaround time of 24 hours with mean reagent cost around $2 per SNP detected.     

DNA from blood samples can be used to genotype patients who have recently received a transfusion

BACKGROUND: The use of hemagglutination to phenotype red cells from recently transfused patients or of red cells that are coated with IgG can be time-consuming and difficult to interpret. Because the molecular bases of many blood group antigens are known, it was investigated whether polymerase chain reaction (PCR) analysis of DNA, from white cells in blood from transfused patients, could be used to predict the blood group antigen profile of a patient. STUDY DESIGN AND METHODS: To prevent problems arising from potentially poor-quality DNA in clinical samples, primers that flanked the polymorphism of interest and that replicated a relatively short PCR amplicon were used. The PCR products, with or without digestion with the appropriate restriction enzyme, were analyzed on gels. Samples were collected from 60 patients who had received from 2 to 50 units of RBCs in the 7 days before sample collection. RBCs from some of these patients were coated with IgG. Analyses for RHD/non-D, RHE/RHe, KEL1/KEL2, FYA/FYB, FY-GATA, JKA/JKB, and GYPA M/N were performed by using assays that had been validated with DNA prepared from untransfused volunteers of known phenotype. The genotyping assays were performed without knowledge of the expected result. RESULTS: The predicted genotype after analysis of the 60 patient samples was that expected from the results of phenotyping. In all cases, the molecular analysis gave a single result; no evidence of chimerism was obtained. CONCLUSION: In each case, the molecular genotype results were in agreement with the blood group antigen as determined by historical phenotyping, phenotyping after hypotonic washing, detection of alloantibodies in the patient's serum, or elution of alloantibody(ies). Under the conditions of these assays, reliable determination of a blood group allele can be made by PCR-restriction fragment length polymorphism testing.     

The A1B genotype expressed as A2B on the red cells of individuals with strong B gene-specific transferases. Results from two paternity cases

Two paternity cases involving black persons are reported in which possible paternal exclusions existed if the A2B red cell phenotype represented the genotype A2B. ABH transferase assays were performed to determine if the A2B phenotypes arose from the genotype A1B or A2B. These assays confirmed that all group A members carried the A1 gene and not the A2 gene. Therefore, the alleged father was not excluded in either case. The B transferase showed increased activity in both AB individuals, possibly accounting for the observed A2B red cell phenotypes. Therefore, in the presence of a B gene, accurate assignment of A subtypes using standard serological methods may not be possible.     

婴儿ABO血型的鉴定及应用于临床输血

为了研究6个月内婴儿ABO血型正确定型方法及影响因素，采用常规血清学法、凝胶柱法及PCR-SSP3种方法对6个月以内的婴儿进行血型鉴定。结果表明：在使用不同方法检测婴儿ABO血型结果不符的33例中，常规血清学方法32例红细胞定型与血清定型不符，1例是由细菌感染产生类B抗原所致的假相合。凝胶柱法可正确定型27例，正确率84.4%。PCR-SSP法可对所有标本做出正确定型。凝胶柱法与PCR-SSP法有显性差异。结论：对6个月以内的婴儿进行ABO血型鉴定，推荐采用凝胶柱技术。凝胶柱法红细胞定型与血清定型相符的婴儿输血采用同型输注，但若婴儿患有感染性疾病时，应考虑到类B抗原引起的假相合。凝胶柱法红细胞定型与血清定型不符的婴儿输血时使用O型洗涤红细胞等成分血，待PCR-SSP法分型结果做出后，改为同型输血。     

天津市滨海新区Rh+无偿献血者Rh血型表型的分布调查

目的 探讨天津市滨海新区Rh+献血者Rh血型表型的分布情况,建立Rh+血型表型数据库,确保临床输血安全,减少输血不良反应的发生.方法 采用简单随机抽样法选择2013年1月至2015年12月天津市第五中心医院输血科留存的2 672份Rh+库存悬浮红细胞为研究对象.研究对象纳入标准:①所有悬液红细胞均来自天津市滨海新区中心血站;②血液经Rh系统中抗-D检测,确定为Rh+血型;③血液入库时,严格核对运输条件、物理外观、血袋封闭及包装、标签等,均应符合血液入库标准.采用微柱凝胶法进行Rh血型系统的抗-D、抗-C、抗-c、抗-E、抗-e检测,并根据抗原检测结果计算Rh血型各表型频率.结果 本组2 672例Rh+无偿献血者的Rh抗原中,按照抗原阳性率由高至低排序,依次为抗-D(100.0％)、抗-e(90.8％)、抗-C(86.7％)、抗-c(58.2％)及抗-E(48.8％)抗原.本组2 672例Rh+无偿献血者中,共检测出9种Rh血型表型,按照抗原频率由高至低排序,依次为CCDee (40.5％)、CcDEe(35.5％)、CcDee(9.4％)、ccDEE(9.2％)、ccDEe(4.1％)血型表型,其他4种表型仅占1.3％(35/2 672).结论 天津市滨海新区Rh+无偿献血人群的Rh血型表型以CCDee为主.建立Rh+血型表型数据库,可为临床及时提供表型相合的Rh+血液,防止由于Rh血型系统不合引起的输血反应,确保临床输血安全.     

红细胞H抗原缺陷型鉴定及其理化功能研究

目的研究红细胞H抗原缺陷型血清学及分子生物学分型特点及其理化功能。方法采用血型血清学鉴定H抗原缺陷型、SSP-PCR进行ABO基因分型,采用红细胞渗透脆性试验、变形性试验检测H抗原缺陷型红细胞理化功能,模拟作为受血者或供血者进行交叉配血试验。结果 6名献血者血样经血清学方法确定为H抗原缺陷型,类孟买基因分型其中3例为h3纯合子,1例为h4纯合子,1例为h1h3杂合子,1例基因分型不确定;H抗原缺陷型红细胞渗透脆性及变形性低于H抗原正常型,H抗原缺陷型作为受血者与A型或B型红细胞主側配血凝集强度强于O型,作为供血者与O型模拟受血者主側配血凝集强度各异。结论 H抗原缺陷型红细胞与抗-H无反应,血清中能检出抗-HI/H,ABO基因正常表达;H抗原缺陷型红细胞渗透脆性及变形性低于H抗原正常型,作为受血者接受O型血液比A型或B型血液安全,作为供血者时,如果交叉配血相合时,可以输给O型受血者。     

Molecular blood typing augments serologic testing and allows for enhanced matching of red blood cells for transfusion in patients with sickle cell disease

BACKGROUND: Sickle cell disease (SCD) patients have dissimilar red blood cell (RBC) phenotypes compared to the primarily Caucasian blood donor base due, in part, to underlying complex Rh and silenced Duffy expression. Gene array–based technology offers high‐throughput antigen typing of blood donors and can identify patients with altered genotypes. The purpose of the study was to ascertain if RBC components drawn from predominantly Caucasian donors could provide highly antigen‐matched products for molecularly typed SCD patients. STUDY DESIGN AND METHODS: SCD patients were genotyped by a molecular array (HEA Beadchip, BioArray Solutions). The extended antigen phenotype (C, c, E, e, K, k, Jk^a, Jk^b, Fy^a, Fy^b, S, s) was used to query the inventory using different matching algorithms; the resulting number of products was recorded. RESULTS: A mean of 96.2 RBC products was available for each patient at basic‐level, 34 at mid‐level, and 16.3 at high‐level stringency. The number of negative antigens correlated negatively with the number of available products. The Duffy silencing mutation in the promoter region (67T>C) (GATA) was found in 96.5% of patients. Allowing Fy(b+) products for patients with GATA increased the number of available products by up to 180%, although it does not ensure prevention of Duffy antibodies in all patients. CONCLUSIONS: This feasibility study provides evidence that centers with primarily Caucasian donors may be able to provide highly antigen‐matched products. Knowledge of the GATA status expands the inventory of antigen‐matched products. Further work is needed to determine the most clinically appropriate match level for SCD patients.     

Genotyping analysis of the MNS blood group system of thalassemia patients with alloantibodies in Iran

BackgroundSerological methods are unreliable for accurate determination of blood group antigens in multi-transfused thalassemia patients. The MNS blood group system has five high-frequency antigens. Many studies demonstrated that some antibodies including anti-S, anti-s, and anti-U may cause acute and delayed transfusion reactions and hemolytic disease of the fetus and newborn. This study aimed to determine the genotype of the MNS blood group in thalassemia patients with alloantibodies by molecular methods.Material and methodsIn this study, 104 blood samples from thalassemia patients were collected. The blood group phenotype for M, N, S and s antigens was determined by the tube hemagglutination method. MNS blood group genotyping was performed using PCR-SSP and DNA Sequencing methods.ResultsAll patients were genotyped with a total of 6 pairs of primers. Discrepancies between genotype and phenotype were observed in 22 patients with S/s alleles and 2 patients with M/N alleles, however, there was full accordance between the results of SSP-PCR and DNA sequencing. The frequency of MNS blood group alleles was determined as follows: 25 % MNSs, 23 % MNss, 21 % MMSs, 9% MMSS, 9% MMss, 8% NNss, 2%MNSS, and NNSS, NNSs, MM genotypes at 1% each.ConclusionIn conclusion, molecular genotyping is more reliable than serological methods in multiple transfusion patients and can lead to a more compatible blood unit for transfusion in these patients.     

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。