title: The Identification of Pompe Disease Mutations in Archival Tissues and Development of a Rapid Molecular-based Test.
abstract: OBJECTIVE
  Pompe disease (glycogen storage disease type II) is a rare autosomal recessive lysosomal storage disease that is caused by acid alpha-glucosidase deficiency. Early enzyme replacement therapy can benefit infants with the disease but the diagnosis is complicated by the rarity of the disease and the heterogeneity of the clinical manifestations. In this study, DNA extracted from archival postmortem formalin-fixed paraffin-embedded tissues was used to identify Pompe disease mutations in Oman and develop a rapid molecular-based test.
  
  
  METHODS
  Intronic primers were designed to amplify short fragments (193-454 base pairs [bp]) from coding exons (2-20) and screen for mutations using direct sequencing (DS).
  
  
  RESULTS
  Two mutations known to cause severe disease were identified in two samples. One was a coding mutation, c.2560C>T (p.Arg854X), and the second was found at a splice acceptor site, c.1327-2A>G. Polymerase chain reaction- and restriction fragment length polymorphism-based tests were designed for the rapid genotyping of the identified mutations.
  
  
  CONCLUSIONS
  These tests can facilitate prenatal diagnosis and help in identifying carriers in families with the identified mutations.
authors.affiliation: Department of Biology, College of Science, Sultan Qaboos University, Muscat, Oman.
authors.forename: Aliya
authors.initials: A
authors.lastname: Alansari
authors.forename: Samira
authors.initials: S
authors.lastname: Al-Rawahi
authors.forename: Taher
authors.initials: T
authors.lastname: Ba-Omar
authors.forename: Mariam
authors.initials: M
authors.lastname: Al-Nabhani
authors.forename: Anand
authors.initials: A
authors.lastname: Date
keywords: Genotyping Techniques; Glucan 1,4-alpha-Glucosidase; Mutations; Oman; Pompe Disease; Tissue