title: Refinement of diagnostic assays for a probable causal mutation for porcine and human malignant hyperthermia.
abstract: The substitutions of T for C1843 in the porcine ryanodine receptor (RYR1) gene, which deletes a HinPI restriction endonuclease site and creates a HgiAI site, and of T for C1840 in human RYR1, which deletes a RsaI site, lead to Cys for Arg substitutions in the ryanodine receptors and are probable causal mutations for malignant hyperthermia (MH). To improve the restriction endonuclease assay of these sites, thereby providing an accurate, reliable diagnosis for MH, introns flanking the exon containing the mutation were sequenced, permitting identification and PCR amplification of a 659-bp porcine gene sequence that contains both constant and variant HgiAI sites and a 922-bp human gene sequence that contains both constant and variant RsaI sites. As a result, these PCR-amplified sequences contain constant internal controls for the reliable differentiation by restriction endonuclease digestion of normal, heterozygous, and MH genotypes.
authors.affiliation: Banting and Best Department of Medical Research, Charles H. Best Institute, University of Toronto, Ontario, Canada.
authors.forename: K
authors.initials: K
authors.lastname: Otsu
authors.forename: M S
authors.initials: MS
authors.lastname: Phillips
authors.forename: V K
authors.initials: VK
authors.lastname: Khanna
authors.forename: S
authors.initials: S
authors.lastname: de Leon
authors.forename: D H
authors.initials: DH
authors.lastname: MacLennan
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis; D005091:Exons; D006801:Humans; D007438:Introns; D008305:Malignant Hyperthermia / Q000175:diagnosis* / Q000235:genetics* / Q000662:veterinary; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D011950:Receptors, Cholinergic / Q000235:genetics; D019837:Ryanodine Receptor Calcium Release Channel; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000235:genetics
chemical_list: D011950:Receptors, Cholinergic; D019837:Ryanodine Receptor Calcium Release Channel; D004247:DNA