{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T00:00:34Z","timestamp":1778803234595,"version":"3.51.4"},"reference-count":22,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T00:00:00Z","timestamp":1745193600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion Medicine"],"published-print":{"date-parts":[[2025,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Mixed agglutination is a serological pattern in some ambiguous ABO blood type identification. This study focused on the serological and molecular genetic characteristics of a B3 phenotype induced by a c.259G\u2009&gt;\u2009T mutation in the <jats:italic>ABO<\/jats:italic> gene.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design and Methods<\/jats:title><jats:p>Serological methods such as gel cards and tubes were used to identify the ABO blood type of the patient, with fluorescent PCR for ABO genotyping and Sanger sequencing for analysing the <jats:italic>ABO<\/jats:italic> exons. Protein 3D Structure was simulated and further analysed using SWISS\u2010MODLE and PyMOL. Both the wild\u2010type (VAL\u201087, <jats:italic>ABO*B.01<\/jats:italic>) and the mutant (p.Val87Leu) plasmids were transfected into Hela cells to assess the agglutination intensity of the transfected cells with anti\u2010B antibodies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Serological testing showed weak expression of the B antigen and mixed agglutination with anti\u2010B antibodies. ABO genotyping indicated the presence of a B allele, but exon sequencing revealed an additional c.259G\u2009&gt;\u2009T mutation in exon 6 based on the <jats:italic>ABO*B.01<\/jats:italic> allele. The simulated three\u2010dimensional structures of the proteins showed increased steric hindrance with mutations, leading to a relatively loose structure. The transfected Hela cells with the mutant (p.Val87Leu) plasmid exhibited a significantly reduced agglutination intensity with anti\u2010B antibodies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Based on comprehensive serological, genetic, and simulation analyses, it is concluded that the c.259G\u2009&gt;\u2009T mutation in exon 6 of the <jats:italic>ABO*B.01<\/jats:italic> allele results in an amino acid change within the enzymatic active site. This alteration likely impacts protein stability and reduces B antigen expression, leading to the B3 subtype phenotype.<\/jats:p><\/jats:sec>","DOI":"10.1111\/tme.13140","type":"journal-article","created":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T21:35:08Z","timestamp":1745271308000},"page":"352-358","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Serological and genetic analysis of a <scp>B3<\/scp> phenotype caused by c.<scp>259G<\/scp>\u2009&gt;\u2009T in the <scp><i>ABO<\/i><\/scp> gene"],"prefix":"10.1111","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-2841-2612","authenticated-orcid":false,"given":"Min","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Transfusion Medicine Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University)  Changsha China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7900-7042","authenticated-orcid":false,"given":"Hongjun","family":"Gao","sequence":"additional","affiliation":[{"name":"Blood Transfusion Institute Jiangsu Zojiwat Biopharmaceutical Co., Ltd.  Wuxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Jiang","sequence":"additional","affiliation":[{"name":"Emergency Research Institute Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University)  Changsha China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fengxia","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Transfusion Medicine The Third Xiangya Hospital of Central South University  Changsha China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xisha","family":"Huan","sequence":"additional","affiliation":[{"name":"Department of Transfusion Medicine Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University)  Changsha China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhen","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Transfusion Medicine Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University)  Changsha China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fen","family":"Yuan","sequence":"additional","affiliation":[{"name":"Department of Transfusion Medicine Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University)  Changsha China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ping","family":"Lei","sequence":"additional","affiliation":[{"name":"Department of Transfusion Medicine Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University)  Changsha China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,4,21]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.12968\/bjon.2023.32.7.326"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/345229a0"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.0042-9007.2001.00134.x"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1159\/000536556"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1011014307683"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1537-2995.2007.01203.x"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-10-387167"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/trf.14209"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/trf.12461"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/trf.17109"},{"issue":"1","key":"e_1_2_10_12_1","first-page":"106","article-title":"11. 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