Detection of non-DeltaGT NCF-1 mutations in chronic granulomatous disease.
OBJECTIVE
Chronic granulomatous disease (CGD) is a rare inherited disorder caused by mutations in the subunits of the NADPH oxidase complex, leaving phagocytes unable to produce superoxide and thereby unable to kill invading microorganisms. A subgroup of CGD patients (approximately 20%) is reported to have mutations in NCF-1 encoding p47-phox, which is part of the cytosolic component of NADPH oxidase. More than 94% of these patients share the same mutation, a 2 bp GT deletion in the GTGT dinucleotide repeat in the start of exon 2. The presence of two pseudogenes more than 98% homologous to the functional NCF-1 has complicated the identification of other mutations in the gene. The aim of this study was to find a general technique for detection of non-GT deletion mutations in the coding region of NCF-1.


RESULTS
A technique involving GeneScan analysis followed by amplification of cDNA with intact dinucleotide repeat was set up and used to identify a novel mutation in exon 7 of NCF-1 in a patient with autosomal recessive CGD, explaining the disease by changing a UGG codon to a premature UGA STOP codon.


CONCLUSIONS
The method is generally applicable for the detection of NCF-1 mutations in patients with suspected CGD.
Jakobsen
Marianne Antonius
MA
Department of Clinical Immunology, Odense University Hospital, Odense C, Denmark. maj@dadlnet.dk
Pedersen
Svend Stenvang
SS
Barington
Torben
T
D000293:Adolescent; D018388:Codon, Terminator; D004252:DNA Mutational Analysis / Q000379:methods*; D005091:Exons; D005260:Female; D005838:Genotype; D006105:Granulomatous Disease, Chronic / Q000235:genetics*; D006801:Humans; D008297:Male; D019255:NADPH Oxidases / Q000235:genetics*; D017354:Point Mutation; D017384:Sequence Deletion
D018388:Codon, Terminator; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1