title: Rapid and cost effective detection of small mutations in the DMD gene by high resolution melting curve analysis.
abstract: Duchenne/Becker muscular dystrophy (DMD/BMD) is caused by large deletions or duplications in two-thirds of the cases. The remaining one-third DMD patients have small mutations in the DMD gene. Screening for such small mutations is a daunting and costly task. High resolution melting curve analysis (HR-MCA) followed by sequencing for amplicons with altered melting profiles can be used to scan DNA for small alterations. We first validated the technique as screening procedure for the DMD gene and then screened a group of unrelated 22 DMD/BMD patients and 11 females. We managed to identify all previously found mutations by means of HR-MCA, which provided its validation. Furthermore, 17 different pathogenic mutations were found in the screening group, of which 10 were novel. Our results provide validation of HR-MCA as a powerful and inexpensive pre-sequencing scanning method. This technology is now ready for routine diagnostic use on DMD/BMD patients and female carriers.
authors.affiliation: Center for Human and Clinical Genetics, LUMC, Einthovenweg 20, Leiden, The Netherlands. r.al_momani.kg@lumc.nl
authors.forename: Rowida
authors.initials: R
authors.lastname: Almomani
authors.forename: Nienke
authors.initials: N
authors.lastname: van der Stoep
authors.forename: Egbert
authors.initials: E
authors.lastname: Bakker
authors.forename: Johan T
authors.initials: JT
authors.lastname: den Dunnen
authors.forename: Martijn H
authors.initials: MH
authors.lastname: Breuning
authors.forename: Ieke B
authors.initials: IB
authors.lastname: Ginjaar
mesh_terms: D000328:Adult; D002648:Child; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods*; D016189:Dystrophin / Q000235:genetics*; D005260:Female; D014644:Genetic Variation; D006579:Heterozygote; D006801:Humans; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction; D013696:Temperature; D013997:Time Factors; D044366:Transition Temperature
chemical_list: C484258:DMD protein, human; D016189:Dystrophin