BACKGROUND: In some cases, a definitive confirmation of dysferlinopathy cannot be achieved by DNA test, because the mutation is detected in one allele only. PATIENTS AND METHODS: DYSFERLIN expression in skeletal muscle and peripheral blood monocytes (PBM) was studied by Western blot in two unrelated adult patients. The comparative C(T) method (ΔΔC(T) ) was used to calculate relative changes in dysferlin mRNA determined from real-time quantitative PCR experiments. The dysferlin gene was studied by direct sequencing of cDNA and genomic DNA and by Multiplex Ligation-dependent Probe Amplification (MLPA) analysis. RESULTS: A comparable severe reduction in dysferlin was demonstrated in both skeletal muscle and PBM. The expression of dysferlin mRNA was significantly reduced. A novel mutation in exon 47 (c.5289G>C) of the dysferlin gene in the heterozygous state, causing an amino acid change (p.Glu1763Asp), was detected in both patients. The MLPA analysis did not reveal any deletion or duplication. CONCLUSIONS: Dysferlin and/or dysferlin mRNA abnormalities are diagnostic for dysferlinopathy when mutational analysis detects a mutation in one allele only. Analysis of dysferlin mRNA can be helpful for distinguishing symptomatic heterozygotes from such patients.
BACKGROUND: In some cases, a definitive confirmation of dysferlinopathy cannot be achieved by DNA test, because the mutation is detected in one allele only. PATIENTS AND METHODS: DYSFERLIN expression in skeletal muscle and peripheral blood monocytes (PBM) was studied by Western blot in two unrelated adult patients. The comparative C(T) method (ΔΔC(T) ) was used to calculate relative changes in dysferlin mRNA determined from real-time quantitative PCR experiments. The dysferlin gene was studied by direct sequencing of cDNA and genomic DNA and by Multiplex Ligation-dependent Probe Amplification (MLPA) analysis. RESULTS: A comparable severe reduction in dysferlin was demonstrated in both skeletal muscle and PBM. The expression of dysferlin mRNA was significantly reduced. A novel mutation in exon 47 (c.5289G>C) of the dysferlin gene in the heterozygous state, causing an amino acid change (p.Glu1763Asp), was detected in both patients. The MLPA analysis did not reveal any deletion or duplication. CONCLUSIONS: Dysferlin and/or dysferlin mRNA abnormalities are diagnostic for dysferlinopathy when mutational analysis detects a mutation in one allele only. Analysis of dysferlin mRNA can be helpful for distinguishing symptomatic heterozygotes from such patients.
Authors: Ines Garcia-Consuegra; Alberto Blázquez; Juan Carlos Rubio; Joaquín Arenas; Alfonsina Ballester-Lopez; Adrián González-Quintana; Antoni L Andreu; Tomàs Pinós; Jaume Coll-Cantí; Alejandro Lucia; Gisela Nogales-Gadea; Miguel A Martín Journal: Genet Med Date: 2016-02-25 Impact factor: 8.822
Authors: Marina Mora; Corrado Angelini; Fabrizia Bignami; Anne-Mary Bodin; Marco Crimi; Jeanne-Hélène Di Donato; Alex Felice; Cécile Jaeger; Veronika Karcagi; Yann LeCam; Stephen Lynn; Marija Meznaric; Maurizio Moggio; Lucia Monaco; Luisa Politano; Manuel Posada de la Paz; Safaa Saker; Peter Schneiderat; Monica Ensini; Barbara Garavaglia; David Gurwitz; Diana Johnson; Francesco Muntoni; Jack Puymirat; Mojgan Reza; Thomas Voit; Chiara Baldo; Franca Dagna Bricarelli; Stefano Goldwurm; Giuseppe Merla; Elena Pegoraro; Alessandra Renieri; Kurt Zatloukal; Mirella Filocamo; Hanns Lochmüller Journal: Eur J Hum Genet Date: 2014-12-24 Impact factor: 4.246