Development of a feasible assay for the detection of GAA mutations in patients with Pompe disease
Journal
Clinica Chimica Acta
Journal Volume
429
Pages
18-25
Date Issued
2014
Author(s)
Abstract
Background: Pompe disease is an inherited autosomal recessive deficiency of acid α-glucosidase (GAA) and is due to pathogenic sequence variants in the corresponding GAA gene. While the analysis of enzyme activity remains the diagnostic test of choice for individuals with Pompe disease, mutation analysis remains for establishing a definitive diagnosis. Methods: High resolution melting (HRM) analysis was performed to screen GAA mutations. Genomic DNA was extracted from peripheral blood samples of the two patients with Pompe disease and 250 normal controls. Exons 2 through 20 of the GAA gene were screened by the HRM analysis. The results were subsequently confirmed by direct sequencing. Results: This assay proved to be feasible in detecting seven known (c.2T>C, c.1726G>A, c.1845G>A, c.1935C>A, c.1958C>A, c.2238G>C, and c.2815_2816del) GAA mutations. Each mutation could be readily and accurately identified in the difference plot curves. We estimated the carrier frequency of the most common mutation, c.1935G>A (p.D645E), in the Taiwanese population to be 0.2%. Conclusions: In clinical practice, we suggest that HRM analysis is assumed as a fast and reliable method for screening GAA gene mutations especially the most common mutations which are responsible for Pompe disease among the Taiwanese populations. ? 2013 Elsevier B.V.
Subjects
Acid α-glucosidase (GAA) gene; High resolution melting; Mutation analysis; Pompe disease
SDGs
Other Subjects
creatine kinase; genomic DNA; glucan 1,4 alpha glucosidase; phosphatase; acid alpha glucosidase gene; article; blood sampling; case report; child; controlled study; DNA extraction; enzyme activity; female; gene frequency; gene mutation; glycogen storage disease type 2; high resolution melting analysis; human; human tissue; male; muscle biopsy; mutational analysis; preschool child; priority journal; real time polymerase chain reaction; school child; sequence analysis; acid α-glucosidase; Acid α-glucosidase (GAA) gene; DBS; denaturing high performance liquid chromatography; DHPLC; dried blood spot; GAA; High resolution melting; high resolution melting; HRM; Mutation analysis; PCR; polymerase chain reaction; Pompe disease; WCN; weighted contact number; alpha-Glucosidases; Base Sequence; Child; Child, Preschool; Conserved Sequence; DNA Mutational Analysis; Feasibility Studies; Female; Gene Frequency; Glycogen Storage Disease Type II; Humans; Male; Nucleic Acid Denaturation; Real-Time Polymerase Chain Reaction; Transition Temperature
Publisher
Elsevier
Type
journal article