title: Rapid molecular diagnosis of genetic diseases by high resolution melting analysis: fabry and glycogen storage 1A diseases.
abstract: For inborn errors of metabolism, high resolution melting analysis (HRMA) is a rapid, efficient, simple, and inexpensive method for mutation/rare variant screening. HRMA is a recent molecular technique for genotyping single-nucleotide polymorphisms without using probes. Here we apply HRMA to the α-galactosidase a (GLA) and glucose-6-phosphatase-alpha (G6PC) genes for mutation detection of patients with Fabry disease (MIM 301500) and glycogen storage disease type 1A (GSD1A; MIM 232200), respectively. To evaluate the procedure, genomic DNAs were blindly tested for known GLA mutations (c.658C>T, c. 679C>T, c.772G>A, c.796G>A, or c.718-719delAA) in three affected males and two obligate heterozygotes with Fabry disease, a G6PC mutation (c.247C>T) in a patient homozygous for that lesion, and 10 healthy control Turkish individuals. HRMA clearly detected the mutant amplicons and discriminated them from all wild-type GLA or G6PC amplicons. HRMA proved to be a sensitive, specific, and cost-effective mutation screening method for the rapid molecular diagnosis of these inborn errors of metabolism, indicating that the technique can be readily adapted to other genetic diseases.
authors.affiliation: 1 Department of Pediatric Metabolic Disorders and Pediatric Genetics, Gazi University Faculty of Medicine , Ankara, Turkey .
authors.forename: Fatih
authors.initials: F
authors.lastname: Ezgu
authors.forename: Yoruk
authors.initials: Y
authors.lastname: Divanoglu
authors.forename: Murat
authors.initials: M
authors.lastname: Polat
authors.forename: Sitkiye
authors.initials: S
authors.lastname: Bahceci
authors.forename: Alev
authors.initials: A
authors.lastname: Hasanoglu
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Desnick
mesh_terms: D016022:Case-Control Studies; D000795:Fabry Disease / Q000175:diagnosis* / Q000235:genetics; D005260:Female; D005953:Glycogen Storage Disease Type I / Q000175:diagnosis* / Q000235:genetics; D006579:Heterozygote; D006801:Humans; D008297:Male; D020125:Mutation, Missense; D057089:Pathology, Molecular; D020641:Polymorphism, Single Nucleotide