Genetic diagnosis of 21-hydroxylase deficiency: DGGE-based mutation scanning of CYP21.
Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is caused by mutations in the gene CYP21 encoding the enzyme steroid 21-hydroxylase. In addition to deletions, approximately 20 different point mutations have been reported, and still novel mutations are detected. This makes genetic diagnosis as well as carrier detection of 21-hydroxylase deficiency a complicated matter. We developed a simple nonradioactive assay based on the polymerase chain reaction (PCR) in combination with denaturing gradient gel electrophoresis (DGGE) to screen for mutations in the CYP21 gene. DGGE allows a fast scanning of PCR-amplified segments of genes for the presence or absence of any single base pair alterations. We have performed this technique on the coding sequence and intron-exon junctions of CYP21. Our results emphasize that this procedure constitutes a fast and reliable approach when performing diagnosis of 21-hydroxylase deficiency.
Ohlsson
G
G
Department of Clinical Genetics, Juliane Marie Center, University Hospital, Rigshospitalet, Copenhagen, Denmark. olsson@biobase.dk
Müller
J
J
Schwartz
M
M
D000312:Adrenal Hyperplasia, Congenital / Q000175:diagnosis*; D019362:Cytochrome P-450 CYP2B1 / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D019362:Cytochrome P-450 CYP2B1