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

Abstract

<b><i>Background:</i></b> Next-generation sequencing (NGS) technology has been recently introduced into blood group genotyping; however, there are few studies using NGS-based blood group genotyping in real-world clinical settings. In this study, we applied NGS-based blood group genotyping into various immunohaematology cases encountered in routine clinical practice. <b><i>Methods:</i></b> This study included 4 immunohaematology cases: ABO subgroup, ABO chimerism, antibody to a high-frequency antigen (HFA), and anti-CD47 interference. We designed a hybridization capture-based NGS panel targeting 39 blood group-related genes and applied it to the 4 cases. <b><i>Results:</i></b> NGS analysis revealed a novel intronic variant (NM_020469.3:c.29-10T&#x3e;G) in a patient with an A_el phenotype and detected a small fraction of <i>ABO</i>*<i>A1.02</i> (approximately 3–6%) coexisting with the major genotype <i>ABO</i>*<i>B.01</i>/<i>O.01.02</i> in dizygotic twins. In addition, NGS analysis found a homozygous stop-gain variant (NM_004827.3:c.376C&#x3e;T, p.Gln126*; <i>ABCG2</i>*<i>01N.01</i>) in a patient with an antibody to an HFA; consequently, this patient’s phenotype was predicted as Jr(a−). Lastly, blood group phenotypes predicted by NGS were concordant with those determined by serology in 2 patients treated with anti-CD47 drugs. <b><i>Conclusion:</i></b> NGS-based blood group genotyping can be used for identifying <i>ABO</i> subgroup alleles, low levels of blood group chimerism, and antibodies to HFAs. Furthermore, it can be applied to extended blood group antigen matching for patients treated with anti-CD47 drugs.