Mutational analysis of patients with p47-phox–deficient chronic granulomatous disease: The significance of recombination events between the p47-phox gene (NCF1) and its highly homologous pseudogenes

Objective The aim of this study was to determine the molecular basis of p47-phox–deficient chronic granulomatous disease (CGD), the most common autosomal recessive form of the disease. CGD is an inherited condition characterized by defective oxygen radical production due to defects in the phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Mutational analysis of p47-phox–deficient CGD patients previously demonstrated that the majority of patients have a GT dinucleotide (ΔGT) deletion at the start of exon 2, a signature sequence also observed in the highly homologous pseudogenes of NCF1. Materials and Methods We performed genetic analysis of NCF1 and its pseudogenes using genomic DNA in 29 p47-phox–deficient CGD patients from 22 separate families. First-strand cDNA analysis was performed in 17 of the 29 patients. Results We confirmed the significance of the ΔGT mutation; in 27 of 29 patients, only the ΔGT sequence was detectable. All but one of the 27 had at least one additional signature sequence, specific to the pseudogene, in either intron 1 and/or intron 2. We extended our analysis to look at signature sequence differences in exons 6 and 9 and detected both the wild-type and pseudogene sequences in all patients tested. Conclusions Although detection of only ΔGT sequence accounts for over 85% of affected patients, the molecular basis is most likely due to partial cross-over events between the wild-type and pseudogene(s) of p47-phox at different recombination sites. Our results suggest that complete gene conversion or deletion of the p47-phox gene (NCF1) occurs rarely, if it all.