High-resolution melting analysis for genotyping Duffy, Kidd and Diego blood group antigens.

High-resolution melting (HRM) analysis is a simpler genotyping method than allele-specific PCR, PCR-restriction fragment length polymorphism and multiplex PCR. Duffy, Kidd and Diego are clinically important blood group antigens. We used a novel method to genotype these three blood group antigens. Purified genomic DNA extracts of blood samples (354 Duffy, 347 Kidd and 457 Diego) were amplified using specific amplification primers. HRM curves were obtained by HRM analysis. Results were in complete concordance with those obtained for previous phenotypes and genotypes. Nucleotide substitutions for these blood group antigens were differentiated by the HRM curves. HRM analysis is a simple genotyping method and is an alternative to serological typing. Our results suggest that HRM analysis can also be used for genotyping blood group antigens whose allotype specificity is determined by single nucleotide substitutions.