Genotyping of 50 RhD variant samples: implication for transfusion ad pregnancy management
10.13303/j.cjbt.issn.1004-549x.2025.12.009
- VernacularTitle:50例RhD变异型的分型研究及输血和孕期管理建议
- Author:
Ke WANG
1
;
Xiaojie MA
2
;
Hailin LI
1
;
Jizhi WEN
3
Author Information
1. Ji'an Central Blood Station, Ji'an 343000, China
2. Blood Transfusion Department, People's Hospital of Xin jiang Uygur Autonomous Region, Urumqi 830001, China
3. Guangzhou Blood Center, Guangzhou 510095, China; Institute of Blood Transfusion and Hematology, Guangzhou Medical University, Guangzhou 510180, China; The Key Medical Laboratory of Guangzhou, Guangzhou 510095, China
- Publication Type:Journal Article
- Keywords:
RhD variant;
RHDgenotyping;
multiple ligation-dependent probe amplification (MLPA);
Sanger sequencing
- From:
Chinese Journal of Blood Transfusion
2025;38(12):1707-1712
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To genotype 50 RhD variant samples from Guangzhou, China, using our previously established genotyping strategy, thereby providing guidance for transfusion management and antenatal monitoring in RhD-variant individuals. Methods: Between June and August 2024, fifty samples identified as RhD variants during RhD-negative confirmation testing at Guangzhou Blood Center were collected. Serological testing for the D antigen was performed with two different anti-D reagents, and the epitope profiles of the D antigen were determined using a commercial panel of monoclonal anti-D reagents containing nine kinds of monoclonal anti-D. Genomic DNA was extracted, and high-resolution melting (HRM) analysis was applied to detect the Asian-type DEL (RHD
1227A). Subsequently, RHD genotyping was carried out using Multiplex Ligation-dependent Probe Amplification (MLPA) and Sanger sequencing. Results: Among the 50 D variant samples, 17 (34.0%) Asian type DEL samples were detected by HRM, including 13 cases with RHD
DEL1/01N.01 genotype and 4 cases with RHD
DEL1/DEL1 genotype. Eleven (11/50, 22.0%) samples were typed as DVI by the epitope profiles of D antigen. The epitope profiles of D antigen combined with Sanger sequencing of exon 6 identified 5 (5/50, 10.0%) cases of RHD
weak partial 15/01N.01. MLPA combined with Sanger sequencing identified two cases of RHD
DVI.3/DEL1, representing 4.0% (2/50) of the samples. Additionally, the following RHD genotypes were each detected in one case: RHD
weak D type 18/01N.04, RHD
weak D type 72/01N.01, RHD
weak D type 95/DEL1, RHD
weak D type 114/DEL1, RHD
weak D type 136/DEL1, RHD
weak D type 147/01N.01, RHD
496G/496G, RHD
536C/01N.01, RHD
689A/689A, RHD
689A/DEL1, RHD
DEL32/DEL1, RHD
DV.1/01N.01, RHD
DV.5/01N.01, RHD
01.01/01N.01, and RHD
01/01N.01. Conclusion: Fifty D variant individuals were typed using our previously established serological and molecular approach. These findings provide guidance for precision transfusion therapy in RhD variant patients and inform evidence-based decisions regarding anti-D immunoglobulin prophylaxis for RhD variant pregnant women.
