title: Novel mutations of dystrophin gene in DMD patients detected by rapid scanning in biplex exons DHPLC analysis.
abstract: Mutations in the dystrophin gene result in both Duchenne and Becher muscular dystrophies (DMD and BMD). Approximately 65% of all mutations causing DMD are deletions (60%) or duplications (5%) of large segments of this gene, spanning one exon or more. Due to the large size of the dystrophin gene (79 exons), finding point mutations has been prohibitively expensive and laborious. Recent studies confirm the utility of pre-screening methods, as denaturing high-performance liquid chromatography (DHPLC) analysis in the identification of point mutations in the dystrophin gene, with an increment of mutation detection rate from 65% to more than 92%. Here we suggest an alternative and convenient method of DHPLC analysis in order to find mutations in a more rapid and less expensive way by introducing the analysis of 16 couples of dystrophin amplicons, in biplex exons DHPLC runs. Using this new protocol of biplex exons DHPLC screening, new mutations were identified in four male patients affected by DMD who had tested negative for large DNA rearrangements.
authors.affiliation: Medical Genetics Service, IRCCS Casa Sollievo della Sofferenza Hospital, Poliambulatorio Giovanni Paolo II, Viale Padre Pio, San Giovanni Rotondo (FG), Italy. l.muscarella@operapadrepio.it
authors.forename: Lucia Anna
authors.initials: LA
authors.lastname: Muscarella
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: Piemontese
authors.forename: Raffaela
authors.initials: R
authors.lastname: Barbano
authors.forename: Antonina
authors.initials: A
authors.lastname: Fazio
authors.forename: Vito
authors.initials: V
authors.lastname: Guarnieri
authors.forename: Alessandro
authors.initials: A
authors.lastname: Quattrone
authors.forename: Leopoldo
authors.initials: L
authors.lastname: Zelante
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D016189:Dystrophin / Q000235:genetics*; D005091:Exons / Q000235:genetics*; D020022:Genetic Predisposition to Disease / Q000235:genetics; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics*; D013997:Time Factors
chemical_list: C484258:DMD protein, human; D016189:Dystrophin