Rapid haplotyping of mutations in the Duffy gene using the polymerase chain reaction and sequence‐specific primers

Abstract

The molecular basis for the antigenic variation and red cell expression of the Duffy antigen system has recently been elucidated. We have developed a simple one‐step method for genotyping the single nucleotide polymorphisms in the promoter and exon of the Duffy gene using the polymerase chain reaction and sequence‐specific primers (PCR‐SSP). This method is also capable of haplotyping alleles at the two polymorphisms as being in cis or trans orientation. Twenty‐four serologically typed Caucasoid and Afro‐Caribbean samples were examined to validate the method, with absolute correlation between phenotype and genotype. A further 30 Gamb‐ian samples were genotyped, confirming homozygosity for the FY*null‐FY*B haplotype. Allele, gene and haplotype frequencies were examined in 100 Caucasoid controls. This method permits the rapid genotyping of large numbers of samples and will prove useful as a clinical and research tool.