title: Deletion pattern in the 21-hydroxylase gene detected by polymerase chain reaction.
abstract: In order to detect deletion mutation and/or gene conversion in the 21-hydroxylase (21-OH) gene, we adopted the polymerase chain reaction (PCR) method followed by electrophoresis. Two pairs of synthesized primers, Ta/1b and 2a/2b, each corresponding to the sequence at the 5' portion of the 21-OH gene, were set for PCR. TaqI digestion of amplified DNA from normal individuals using Ta/1b as primers gave the following three fragments: an active 21-OH gene-derived 559 bp fragment, and pseudogene-derived 364 and 195 bp fragments. Of 16 patients with 21-hydroxylase deficiency (21-OHD) studied, 6 (37%) lacked the 559 bp fragment. These 6 patients also lacked both the 331 and 117 bp MvaI fragments of the PCR product which were obtained with the primers 2a/2b, both being derived from the active 21-OH gene. These results indicate that 6 of the 16 patients have either deletion of the 21-OH gene or conversion of the gene to its tandemly located pseudogene. The method described here provides a rapid diagnosis of 21-OHD.
authors.affiliation: Department of Pediatrics, Nagasaki University School of Medicine, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Kinoshita
authors.forename: T
authors.initials: T
authors.lastname: Matsumoto
authors.forename: T
authors.initials: T
authors.lastname: Kondoh
authors.forename: M
authors.initials: M
authors.lastname: Yoshimoto
authors.forename: N
authors.initials: N
authors.lastname: Niikawa
authors.forename: Y
authors.initials: Y
authors.lastname: Tsuji
mesh_terms: D000312:Adrenal Hyperplasia, Congenital / Q000175:diagnosis / Q000235:genetics; D001483:Base Sequence; D002872:Chromosome Deletion; D002885:Chromosomes, Human, Pair 16; D004247:DNA / Q000032:analysis*; D005260:Female; D005785:Gene Conversion; D005796:Genes / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D011296:Prenatal Diagnosis / Q000379:methods*; D011544:Pseudogenes; D013255:Steroid 21-Hydroxylase / Q000235:genetics*
chemical_list: D009841:Oligonucleotides; D004247:DNA; D013255:Steroid 21-Hydroxylase