title: Screening of the entire ryanodine receptor type 1 coding region for sequence variants associated with malignant hyperthermia susceptibility in the north american population.
abstract: BACKGROUND
  Malignant hyperthermia (MH) is a life-threatening and frequently fatal disorder triggered by commonly used anesthetics. MH susceptibility is a genetically determined predisposition to the development of MH. Mutations in the ryanodine receptor type 1 (RYR1) gene are the major cause of MH susceptibility. The authors sought to develop a reliable genetic screening strategy based on efficient and relatively inexpensive mutation-detection procedures.
  
  
  METHODS
  A cohort (n = 30) of North American MH patients and MH-susceptible individuals was studied. RNA and DNA extracted from muscle tissue or blood lymphocytes were used for analysis. The entire RYR1 coding region was amplified in 57 overlapping fragments and subjected to denaturing high-performance liquid chromatography analysis followed by direct nucleotide sequencing to characterize RYR1 alterations.
  
  
  RESULTS
  Nine previously reported and nine unknown RYR1 mutations were identified in 21 of 30 studied patients (70%). Some of the new mutations were located outside of known mutational "hot spots," suggesting that RYR1 contains previously unknown mutation-prone areas requiring analysis. The North American MH/MH-susceptible population is characterized by a high RYR1 allelic heterogeneity.
  
  
  CONCLUSIONS
  Denaturing high-performance liquid chromatography analysis of RNA samples extracted from the biopsied skeletal muscle followed by DNA sequencing is a highly efficient methodology for RYR1 mutation detection. This approach allows increasing the rate of mutation detection to 70% and identifying mutations in the entire RYR1 coding region.
authors.affiliation: National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892-9404, USA. sambuugn@ninds.noh.gov
authors.forename: Nyamkhishig
authors.initials: N
authors.lastname: Sambuughin
authors.forename: Heather
authors.initials: H
authors.lastname: Holley
authors.forename: Sheila
authors.initials: S
authors.lastname: Muldoon
authors.forename: Barbara W
authors.initials: BW
authors.lastname: Brandom
authors.forename: Astrid M
authors.initials: AM
authors.lastname: de Bantel
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Tobin
authors.forename: Tom E
authors.initials: TE
authors.lastname: Nelson
authors.forename: Lev G
authors.initials: LG
authors.lastname: Goldfarb
mesh_terms: D000818:Animals; D020022:Genetic Predisposition to Disease; D006801:Humans; D008305:Malignant Hyperthermia / Q000235:genetics*; D009154:Mutation; D016366:Open Reading Frames; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics*
chemical_list: D019837:Ryanodine Receptor Calcium Release Channel