In:
Clinical Chemistry, Oxford University Press (OUP), Vol. 48, No. 6 ( 2002-06-01), p. 818-825
Abstract:
Background: Congenital adrenal hyperplasia (CAH) is a frequent autosomal recessive disease, with a wide range of clinical manifestations, most commonly attributable to mutations in the 21-hydroxylase gene (CYP21). Large gene deletions, large gene conversions, a small 8-basepair deletion, and eight point mutations in CYP21 account for ∼95% of all enzyme deficiencies. We developed a new strategy for a rapid CYP21 analysis. Methods: DNA samples from 40 CAH patients previously genotyped by direct DNA sequencing were reanalyzed by allele-specific amplification of the functional CYP21 gene followed by a multiplex minisequencing reaction using 13 primers. In addition, a second PCR that amplified a part of exon 3 was used to demonstrate the presence or absence of at least one functional gene. Results: The assay detected the P453S mutation and nine of the most common mutations (P30L, intron 2 splice, Δ8bp, I172N, exon 6 cluster, V281L, F306+t, Q318X, and R356W) caused by microconversions from the CYP21P pseudogene. The concordance was 100% for detecting these mutations, including gene deletions and large gene conversions. The 40 patient DNA samples were analyzed in 1.5 working days by one technician (actual hands-on time, 3.5 h). The material cost for analyzing one sample was approximately €10.00 (US $9.00). Conclusions: This novel mutation screening strategy rapidly detects 90–95% of all mutations associated with CAH and appears applicable as a tool for confirmation of increased 17-hydroxyprogesterone found in neonatal CAH screening.
Type of Medium:
Online Resource
ISSN:
0009-9147
,
1530-8561
DOI:
10.1093/clinchem/48.6.818
Language:
English
Publisher:
Oxford University Press (OUP)
Publication Date:
2002
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