Research on the molecular mechanisms of ABO subtypes based on first-generation and third-generation sequencing technologies
10.13303/j.cjbt.issn.1004-549x.2025.07.010
- VernacularTitle:结合一代与三代测序技术分析ABO正反定型不符血型标本的策略研究
- Author:
Chengyan GAO
1
;
Hui ZHANG
2
;
Hang LEI
1
;
Can LOU
1
;
Xiaohong CAI
1
Author Information
1. Department of Transfusion Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
2. The Central Hospital of Minhang District, Shanghai 201100, China
- Publication Type:Journal Article
- Keywords:
ABO subtybes;
first-generation (Sanger) sequencing;
third-generation (long-read) sequencing;
genetic mutation;
intron mutation
- From:
Chinese Journal of Blood Transfusion
2025;38(7):928-933
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To accurately determine the ABO blood group of samples exhibiting forward/reverse grouping discrepancies by combining first-generation (Sanger) and third-generation (long-read) sequencing technologies. Methods: Five samples with ABO forward/reverse grouping discrepancies were selected. Serological testing was conducted using automated blood typing instruments and the tube method. Genotyping was conducted using both Sanger and long-read sequencing technologies. Results: Sanger sequencing identified specific genetic mutations in two samples, with genotypes of ABO
BA. 04/ABO
O.01.01 and ABO
B3.05/ABO
O.01.02. Further analysis with long-read sequencing revealed specific mutations in the +5.8kb region of intron 1 (c.28+5885C>T and c.28+5861T>G) in three samples where mutations were not detected by Sanger sequencing. These mutations affect the expression of the ABO antigens and are likely responsible for the ABO subgroup phenotypes. Conclusion: The integration of Sanger and long-read sequencing technologies effectively identifies genetic variations causing ABO subtypes, providing a scientific basis for enhancing clinical transfusion safety and ensuring accurate blood group determination.
