title: High-resolution melting curve (HRM) analysis to establish CYP21A2 mutations converted from the CYP21A1P in congenital adrenal hyperplasia.
abstract: BACKGROUND
  Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease of an inborn error of steroid metabolism in humans. More than 90% of CAH cases are caused by mutations of the steroid 21-hydroxylase (CYP21A2) gene, and approximately 75% of the defective CYP21A2 genes are generated through an intergenic recombination with the neighboring CYP21A1P pseudogene.
  
  
  METHODS
  A high-resolution melting (HRM) curve analysis was designed to characterize 11 mutation sites of the CYP21A2 gene that commonly appeared in 21-hydroxylase deficiency. Among these 11 mutations, 9 were found in CAH patients, and 2 were mutations created from normal individuals.
  
  
  RESULTS
  From the HRM analysis using 6 fragments of amplicons, we have successfully identified these 11 common disease-causing mutations of the CYP21A2 gene, among which 3 showed 3 distinguishable melting plots; the heteroduplexes showed an upcurved plot, a horizontal plot of homoduplexes of wild-type (WT), and a downcurved plot of homoduplexes of compound mutations.
  
  
  CONCLUSIONS
  The HRM analysis is a 1-step of non-gel resolution technique which saves time and is a low-cost method to undertake such a program for screening CAH patients with the 21-hydroxylase deficiency caused by intergenic conversions from the neighboring CYP21A1P pseudogene.
authors.affiliation: Institute of Clinical Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
authors.forename: Yi-Ching
authors.initials: YC
authors.lastname: Lin
authors.forename: Yu-Chih
authors.initials: YC
authors.lastname: Lin
authors.forename: Ta-Chih
authors.initials: TC
authors.lastname: Liu
authors.forename: Jan-Gowth
authors.initials: JG
authors.lastname: Chang
authors.forename: Hsien-Hsiung
authors.initials: HH
authors.lastname: Lee
mesh_terms: D000312:Adrenal Hyperplasia, Congenital / Q000201:enzymology / Q000235:genetics*; D000483:Alleles; D004247:DNA / Q000235:genetics; D006801:Humans; D009154:Mutation; D009691:Nucleic Acid Denaturation / Q000235:genetics*; D011544:Pseudogenes / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D012984:Software; D013255:Steroid 21-Hydroxylase / Q000235:genetics*; D044366:Transition Temperature
chemical_list: D004247:DNA; C531510:CYP21A2 protein, human; D013255:Steroid 21-Hydroxylase