Assay for propensity for canine malignant hyperthermia

Abstract

A method of determining whether a canine is susceptible to canine malignant hyperthermia is disclosed. In one embodiment, this method comprises the step of obtaining a nucleic acid sample from a canine and examining the DNA sample for the presence or absence of a T1640C mutation, wherein the presence of the mutation indicates that the canine is susceptible to canine malignant hyperthermia.

We claim:  
     
         1 . A method of determining whether a canine is susceptible to canine malignant hyperthermia, comprising the step of obtaining a nucleic acid sample from a canine and examining the sample for the presence or absence of a T1640C mutation, wherein the presence of the mutation indicates that the canine is susceptible to canine malignant hyperthermia.  
     
     
         2 . The method of  claim 1  wherein the nucleic acid sample is a genomic DNA sample.  
     
     
         3 . The method of  claim 2  wherein the DNA is obtained from cheek cells.  
     
     
         4 . The method of  claim 2  wherein the DNA is obtained from muscle cells.  
     
     
         5 . The method of  claim 2  wherein the DNA is obtained from blood cells.  
     
     
         6 . The method of  claim 1  wherein the step of examining the sample for the presence or absence of a T1640C mutation comprises amplifying a portion of the canine RYR1 gene, wherein the portion comprises nucleotide 1640, and examining the amplified product for the presence of the T1640C mutation.  
     
     
         7 . The method of  claim 6  wherein the portion comprises the sequence between Exon 14 and Exon 16 of the RYRI gene.  
     
     
         8 . The method of  claim 6  further comprising the step of digesting the amplification product with a restriction endonuclease.  
     
     
         9 . The method of  claim 7  further comprising the step of digesting the amplified product with a restriction endonuclease.  
     
     
         10 . The method of  claim 8  wherein the restriction endonuclease is Mscl.  
     
     
         11 . The method of  claim 9  wherein the restriction endonuclease is Mscl.  
     
     
         12 . A kit for determining whether a canine is susceptible to canine malignant hyperthermia comprising: 
 (a) a set of primers useful for amplifying at least a portion of the RYRI gene, wherein the amplified portion comprises nucleotide 1640; and    (b) a restriction endonuclease capable of differential cleavage in the presence or absence of a T1640C mutation.    
     
     
         13 . The kit of  claim 12  wherein the restriction endonuclease is Mscl.  
     
     
         14 . The kit of  claim 12  wherein the primers comprise SEQ ID NOs:19 and 20.