title: A novel semiquantitative polymerase chain reaction/enzyme digestion-based method for detection of large scale deletions/conversions of the CYP21 gene and mutation screening in Turkish families with 21-hydroxylase deficiency.
abstract: 21-Hydroxylase deficiency is a recessively inherited disorder resulting from mutations in the CYP21 gene. The CYP21 gene is located along with the CYP21P pseudogene in the human leukocyte antigen major histocompatibility complex region on chromosome 6. Molecular diagnosis is difficult due to the 98% similarity of CYP21 and CYP21P genes and the fact that almost all frequently reported mutations reside on the pseudogene. Allele-specific PCR for the 8 most frequently reported point mutations was performed in 31 Turkish families with at least a single 21-hydroxylase-deficient individual. The allele frequencies of the point mutations were as follows: P30L, 0%; IVS2 (AS,A/C-G,-13), 22.5%; G110delta8nt, 3.2%; I172N, 11.4%; exon 6 cluster (I236N, V237E, M239K), 3.2%; V281L, 0%; Q318X, 8%; and R356W, 9.6%. Large deletions and gene conversions were detected by Southern blot analysis, and the allele frequencies were 9.6% and 22.5%, respectively. Sequence analysis of the gene, performed on patients with only 1 mutated allele, revealed 2 missense mutations (R339H and P435S). A novel semiquantitative PCR/enzyme digestion-based method for the detection of large scale deletions/conversions of the gene was developed for routine diagnostic purposes, and its accuracy was shown by comparison with the results of Southern blot analysis.
authors.affiliation: Department of Pediatrics, Istanbul Medical Faculty, Istanbul University, 34390 Istanbul, Turkey. turgut.tukel@mssm.edu
authors.forename: T
authors.initials: T
authors.lastname: Tukel
authors.forename: O
authors.initials: O
authors.lastname: Uyguner
authors.forename: J Q
authors.initials: JQ
authors.lastname: Wei
authors.forename: M
authors.initials: M
authors.lastname: Yuksel-Apak
authors.forename: N
authors.initials: N
authors.lastname: Saka
authors.forename: D X
authors.initials: DX
authors.lastname: Song
authors.forename: H
authors.initials: H
authors.lastname: Kayserili
authors.forename: F
authors.initials: F
authors.lastname: Bas
authors.forename: H
authors.initials: H
authors.lastname: Gunoz
authors.forename: R C
authors.initials: RC
authors.lastname: Wilson
authors.forename: M I
authors.initials: MI
authors.lastname: New
authors.forename: B
authors.initials: B
authors.lastname: Wollnik
mesh_terms: D000312:Adrenal Hyperplasia, Congenital / Q000235:genetics*; D000483:Alleles; D015139:Blotting, Southern; D005785:Gene Conversion; D017353:Gene Deletion; D005787:Gene Frequency; D006801:Humans; D020125:Mutation, Missense; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D013255:Steroid 21-Hydroxylase / Q000235:genetics*; D019914:Taq Polymerase; D014421:Turkey
chemical_list: D013255:Steroid 21-Hydroxylase; D019914:Taq Polymerase