Objective  To explore the distribution of RhCE phenotypes and the molecular genetic background of D variants among preliminary screening RhD-negative blood donors.

Methods  Blood samples sent for testing to the Linyi Central Blood Station from July 2022 to June 2023 were collected. The Rh phenotype was detected using the test tube method, and RhD blood type confirmation was performed using a broad-spectrum anti-human globulin card (microcolumn method). For samples serologically confirmed as RhD variant, the RHD gene was tested using PCR-SSP. Sequencing analysis was performed on samples with positive amplification of all or part of the exons.

Results  A total of 564 cases were initially screened as RhD negative. The ABO blood type distribution was as follows: O type (32.98%), A type (27.48%), B type (26.06%), and AB type (13.48%). The confirmation results for RhD negative status showed 548 cases (97.16%) as RhD negative and 16 cases (2.84%) as RhD variants. The distribution of Rh phenotypes was ccee, Ccee, ccEe, CCee, CcEe and CCEe. The RhCE phenotypes of RHD variants all contained C or E antigens and there were several alleles which were previously reported. To be specific, DEL RHD1227A homozygous, weak D15, weak D15/DEL RHD1227A, DVI III, RHD-CE(5)-D and RHD-CE(5)-D/711delC were detected in 2 cases, 6 cases, 3 cases, 1 case, 3 cases and 1 case respectively.

Conclusion  The most common RhCE phenotype of RhD negative donors were ccee and Ccee. There was polymorphism in RHD gene with D variant, weak D15 and DEL RHD1227A were the most common. The RhCE phenotype of D variant was associated with RHD genotype.

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