Application of Blood Group Genotyping by Next-Generation Sequencing in Various Immunohaematology Cases

Kim, Tae Yeul; Yu, HongBi; Phan, Minh‐Trang Thi; Jang, Ja‐Hyun; Cho, Duck

doi:10.1159/000517565

Cited by 6 publications

(6 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…A universally applicable method is to transfuse phenotypically and/or genotypically matched RBCs. This approach can be helpful for preventing alloimmunization, leading to potential hemolytic transfusion reactions, which is well demonstrated in cases of safe transfusion in patients on evorpacept using prior genotypic blood group data via next-generation sequencing [ 35 ]. Although there can be debate on the appropriate extent of the type of antigens that need to be matched before RBC transfusion, this approach is particularly useful when serologic methods have limitations in resolving the problem.…”

Section: Discussionmentioning

confidence: 99%

Evorpacept-Induced Interference and Application of a Novel Mitigation Agent, Evo-NR, in Pretransfusion Testing

Yoon,

Kim,

Kim

et al. 2023

Transfus Med Hemother

View full text Add to dashboard Cite

Introduction: Evorpacept is a CD47-blocking agent currently being developed for the treatment of various cancers. Interference by evorpacept in pretransfusion compatibility testing has been reported at limited plasma concentrations. Although various mitigation strategies have been proposed, none are practical. This in vitro study assessed evorpacept-induced interference at extended concentrations and investigated the capability of a novel mitigation agent, Evo-NR. Methods: Antibody screening tests were performed on evorpacept-spiked plasma with (anti-E and anti-Jka) or without alloantibodies at evorpacept concentrations up to 2,000 μg/mL using manual gel cards and automated analyzers. Evorpacept-coated red blood cells (RBCs) (rr [ce/ce], Fy[a+b−], S−s+) were tested by direct antiglobulin testing (DAT) and antigen typing using anti-Fyb and anti-S reagents at indirect antiglobulin testing (IAT) phase. Evo-NR was used to resolve the interference in plasma and RBC samples. Flow cytometry was used to assess the mitigation effects. Results: Evorpacept-spiked plasma showed panreactive interference in antibody screening tests using manual gel cards (2+ to 3+) and automated analyzers (4+). A carryover effect was also observed in the automated analyzers. The use of a 3- to 6-fold molar excess of Evo-NR effectively resolved the interference in the plasma and enabled accurate alloantibody identification. Although the reduction in evorpacept binding to RBCs was identified via flow cytometry, Evo-NR was incapable of resolving the serologic interference observed in DAT and antigen typing at IAT phase. Discussion: Evorpacept showed constant panreactivity and a carryover effect at high concentrations. Evo-NR successfully resolved the interference in the plasma samples and could be considered a practical and efficient mitigation solution. Implementation of Evo-NR has the potential to support RBC transfusion for patients undergoing evorpacept treatment.

show abstract

Section: Discussionmentioning

confidence: 99%

Evorpacept-Induced Interference and Application of a Novel Mitigation Agent, Evo-NR, in Pretransfusion Testing

Yoon,

Kim,

Kim

et al. 2023

Transfus Med Hemother

View full text Add to dashboard Cite

show abstract

“…그런데, 일부 ABO 아형은 ABO 유전형 검사로 그 유전적 원인이 규 명되지 않았다. 이는 ABO 유전자 중 엑손 6과 7 만을 주로 분석했기 때문으로 판단되어, 최근 근 접 프로모터 및 인트론 1 ＋5.8 kb 부위의 변이를 추가로 분석하여 기존 검사의 한계를 일부 극복 하고 있다 [5][6][7][8][9][10][11].…”

Section: 서 론unclassified

“…국내 한 의료기관에서 ABO 유전형 검사의 경 험을 보고한 지 10년이 지났다 [12]. 그 후 ABO 불일치의 원인 규명도 다양해지고, ABO 유전자 분석 범위도 확장되었으며, 차세대염기서열분석 (Next generation sequencing, NGS) 같은 새로운 기 법도 적용되고 있다 [11]. 그런데 ABO discrepancy due to unexpected antibodies comprised 5 cases with autoantibodies, 5 cases with anti-M, 4 cases with anti-P1, 4 cases with unidentified alloantibodies, 1 case with a cold autoantibody, 1 case with an anti-N, and 1 case with a coexisting anti-P1 and cold autoantibody.…”

Section: 서 론unclassified

Clinical Application of ABO Genotyping: 5-Year Experience from a Single Tertiary Care Center in Korea

Lee

Jeong

Seo

et al. 2023

Korean J Blood Transfus

Self Cite

View full text Add to dashboard Cite

Background: ABO genotyping is performed when the exact ABO blood type cannot be determined through serological testing. Conventionally, only exons 6 and 7 of the ABO gene have been analyzed, but our laboratory introduced additional analysis of the proximal promoter and intron 1 ＋5.8 kb site. Accordingly, we report the clinical use of ABO genotyping and distribution of the ABO subgroups based on our experience over the past 5 years. Methods: A total of 265 samples tested at the Samsung Medical Center from August 2017 to July 2022 were retrospectively analyzed at their request. Serological ABO blood typing and direct sequencing of exons 6 and 7 were performed on all samples, and additional analysis of the regulatory region of the ABO gene was performed on 17 samples. Since some of the ABO discrepant cases revealed multiple causes, a total of 339 causes among 238 ABO discrepant cases were analyzed. Results: Among the total of 265 samples, 89.8% (238/265) exhibited ABO discrepancies. Weak red cell reactivity (51.6%, 175/339) was the most common cause of ABO discrepancy, followed by extra serum reactivity (35.7%, 121/339). Among the samples, 40.8% (108/265) were identified as ABO subgroups. Among the 108 ABO subgroup alleles, cisAB.01 in exons 6 and 7 accounted for 82 cases (75.9%, 82/108), and two g.10925C＞T mutations in intron 1 ＋5.8 kb were identified. Conclusion: Through our recent experience of the last 5 years of ABO genotyping, we elucidated the cause of ABO discrepancies and ABO subgroup alleles. The extended sequencing of the regulatory region of the ABO gene was helpful for further understanding the ABO discrepancy caused by weak red cell reactivity.

show abstract

“…In contrast, next-generation sequencing (NGS)-based assays, particularly wholegenome sequencing (WGS), have the potential to detect all blood group alleles, allowing accurate prediction of null phenotypes. [4][5][6][7] The prevalence of null alleles is essential in determining whether screening in the local population is worthwhile and-if SNV-based molecular assays are usedwhich alleles should be included in population screening; however, data on the Korean population are insufficient. The Korean Reference Genome Database (KRGDB) is a collection of genomic variants found in WGS data of 1722 Korean individuals.…”

Section: Introductionmentioning

confidence: 99%

“…SNV‐based molecular assays can detect all the known null alleles but would have to include a vast number of mutations and be updated regularly, which is not practical. In contrast, next‐generation sequencing (NGS)‐based assays, particularly whole‐genome sequencing (WGS), have the potential to detect all blood group alleles, allowing accurate prediction of null phenotypes 4–7 …”

Section: Introductionmentioning

confidence: 99%

Prevalence and molecular basis of null blood group phenotypes in the Korean population: Analysis using a public database

Kim,

Park,

Jang

et al. 2023

Transfusion

Self Cite

View full text Add to dashboard Cite

BackgroundNull phenotypes are characterized by complete absence of all antigens within a blood group system and caused by null variants (e.g., nonsense, frameshift, initiation codon, and canonical splice site variants) in the genes encoding the antigens. Knowing the prevalence and molecular basis of null phenotypes is essential to establish a rare donor program, and the aim of this study was to reveal the prevalence and molecular basis of null phenotypes using the Korean Reference Genome Database (KRGDB) containing whole‐genome sequences of 1722 Korean individuals.Study Design and MethodsPopulation allele frequencies of null alleles in 39 blood group systems except ABO, MNS, Rh, Lewis, and FORS were obtained from the KRGDB. The prevalence of null phenotypes was calculated using Hardy–Weinberg equation.ResultsThe prevalence of null phenotypes were estimated to be less than 0.001% in all blood group systems except JR and SID. The prevalence of the Jr(a−) and Sd(a−) phenotypes were estimated to be 0.0453% and 0.2323%, respectively. The most frequent null allele of the JR system was ABCG2*01N.01, accounting for approximately 85% of null alleles.DiscussionOur approach using a public database allowed us to investigate the prevalence and molecular basis of null phenotypes in the Korean population, which will serve as a guide for establishing a rare donor program in Korea. Considering the clinical significance, Jr(a−) is an important null phenotype that should be typed in the Korean population, and molecular assays targeting the most frequent allele ABCG2*01N.01 may be useful in detecting this phenotype.

show abstract

Application of Blood Group Genotyping by Next-Generation Sequencing in Various Immunohaematology Cases

Cited by 6 publications

References 43 publications

Evorpacept-Induced Interference and Application of a Novel Mitigation Agent, Evo-NR, in Pretransfusion Testing

Evorpacept-Induced Interference and Application of a Novel Mitigation Agent, Evo-NR, in Pretransfusion Testing

Clinical Application of ABO Genotyping: 5-Year Experience from a Single Tertiary Care Center in Korea

Prevalence and molecular basis of null blood group phenotypes in the Korean population: Analysis using a public database

Contact Info

Product

Resources

About