Sequencing of the ART4 gene in sub-Saharan cohorts reveals ethnic differences and two new DO alleles: DO*B-Ile5Thr and DO*B-Trp266Arg.

BACKGROUND:Given the high heterogeneity of sub-Saharan populations especially between nonpygmoids and pygmoids, differences are expected during investigation of the DO/ART4 gene. STUDY DESIGN AND METHODS:Using genomic DNA extracted from blood samples collected from 77 Tswa pygmoids and 39 Teke and seven San nonpygmoids, DO coding regions were amplified and sequenced. A tetra-primer amplification refractory mutation system-polymerase chain reaction method was developed to specifically detect the DO*B-SH-Gln149Lys variant. Membrane expression of newly identified variant alleles in K562-transduced cells was studied by flow cytometry. RESULTS:Extensive polymorphism was confirmed in Teke or San nonpygmoids and Tswa pygmoids with, respectively, 12, zero, and 24 DO*A; 54, 10, and 115 DO*B or DO*B-WL; five, zero, and 14 DO*HY; and six DO*JO alleles in Teke only. The DO*B-SH-Gln149Lys variant was observed as the third most frequent after the DO*HY and DO*JO alleles. Two novel DO*B alleles were identified in the San samples, that is, DO*B-Ile5Thr and DO*B-Trp266Arg. Study of K562-transduced cells showed that compared to the DO*B allele, DO*B-Ile5Thr was expressed more strongly while DO*B-Trp266Arg variant was expressed to a lesser extent and was not recognized by MIMA-123 monoclonal antibodies. CONCLUSION:Sequencing analysis showed more allelic combinations in nonpygmoids than in pygmoids with high frequencies of DO*HY, DO*JO, and DO*B-SH-Gln149Lys variant alleles. This finding underlines the importance of including DO*HY and DO*JO single-nucleotide polymorphisms in genotyping tests to improve transfusion safety. Characterization of two novel DO*B alleles highlights the value of testing selected ethnic groups in understanding DO allele diversity.

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