High‐resolution Melting Facilitates Mutation Screening of PYGM in Patients with McArdle Disease
Open Access
- 8 March 2009
- journal article
- Published by Wiley in Annals of Human Genetics
- Vol. 73 (3), 292-297
- https://doi.org/10.1111/j.1469-1809.2009.00512.x
Abstract
Mutations in PYGM, encoding the muscle‐specific glycogen phosphorylase (myophosphorylase), are responsible for McArdle disease. Among Caucasians, a large proportion of patients are homozygous for the R50X mutation, but other mutations can affect all the 20 exons of PYGM, making mutation detection laborious. We have developed a high‐resolution melting (HRM) assay for mutation detection in PYGM. Twelve McArdle patients were investigated, in whom pre‐screening had ruled out homozygosity or compound heterozygosity for the two common G205S and R50X mutations. In total, we identified 16 different variations. Thirteen of these are pathogenic, and three were classified as polymorphisms. Nine variations had not previously been described. One of the novel mutations, c.2430C > T, was initially predicted to result in a silent G810G change, but cDNA analysis demonstrated that the mutation led to abnormal mRNA processing. The HRM protocol reduced the need for direct sequencing by approximately 85%, and is a good approach to search for new mutations in PYGM.Keywords
This publication has 31 references indexed in Scilit:
- Validation of High-Resolution DNA Melting Analysis for Mutation Scanning of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) GeneThe Journal of Molecular Diagnostics, 2008
- Evaluation of in silico splice tools for decision-making in molecular diagnosisHuman Mutation, 2008
- High resolution melting applications for clinical laboratory medicineExperimental and Molecular Pathology, 2008
- Genotype modulators of clinical severity in McArdle diseaseNeuroscience Letters, 2007
- A proposed molecular diagnostic flowchart for myophosphorylase deficiency (McArdle disease) in blood samples from Spanish patientsHuman Mutation, 2007
- Variable presentation of the clinical phenotype of McArdle's disease in a kindred harbouring a novel compound genotype in the muscle glycogen phosphorylase geneNeuroscience Letters, 2005
- A diagnostic cycle test for McArdle's diseaseAnnals of Neurology, 2003
- Two novel mutations in the myophosphorylase gene in a patient with McArdle diseaseMuscle & Nerve, 2002
- Listening to silence and understanding nonsense: exonic mutations that affect splicingNature Reviews Genetics, 2002
- Improved Splice Site Detection in GenieJournal of Computational Biology, 1997