Genotyping Assays for the Canine Degenerative Myelopathy-Associated c.118G>A (p.E40K) Mutation of the <i>SOD1</i> Gene Using Conventional and Real-Time PCR Methods: A High Prevalence in the Pembroke Welsh Corgi Breed in Japan

Abstract

ABSTRACT. Canine degenerative myelopathy is an adult-onset, progressive neurodegenerative disease that occurs in multiple dog breeds, particularly Pembroke Welsh Corgis. Recently, a degenerative myelopathy-associated mutation of the canine SOD1 gene was identified as c.118G>A (p.E40K). In the present study, genotyping assays using conventional and real-time PCR methods were developed, and a preliminary genotyping survey was performed on 122 randomly selected Pembroke Welsh Corgis without any degenerative myelopathy-related clinical signs to determine the current allele frequency in Japan. Both of the assays provided clear-cut genotyping. The survey demonstrated the frequencies of the G/G wild-type, G/A heterozygote and A/A homozygote to be 9.0, 42.6 and 48.4%, respectively, indicating that the prevalence of the mutant A allele (69.7%) in Pembroke Welsh Corgis is extremely high in Japan.

A retrospective study of the prevalence of the canine degenerative myelopathy associated superoxide dismutase 1 mutation (SOD1:c.118G > A) in a referral population of German Shepherd dogs from the UK

“…The SOD1:c.118G > A mutation, associated with CDM, was originally identified in a genome-wide association study and characterised by sequence-based typing [ 13 ]. Recent studies have used conventional and real-time PCR techniques to develop more rapid and cost-effective methods of genotyping dogs for this particular mutation [ 14 – 16 ]. The present study employed PCR followed by RFLP analysis, utilising the Eco571 enzyme, demonstrating that this technique allows rapid and simple genotyping dogs for the SOD1:c.118G > A mutation.…”

Accumulation and aggregate formation of mutant superoxide dismutase 1 in canine degenerative myelopathy

“…All dogs with A/A or A/G genotypes were PWC, while the three other dogs carrying G/G were two Beagles and a Maltese. The genotypes of the dogs used in the present study were determined using a previously reported real-time PCR method (Chang et al, 2013). Formalin-fixed paraffin-embedded sections of the caudal thoracic spinal cord were prepared from each dog.…”

Canine degenerative myelopathy: a model of human amyotrophic lateral sclerosis

“…In a recent study (Chang et al, 2013), genotyping assays using conventional and real-time PCR methods were developed, and a preliminary genotyping survey was performed on 122 randomly selected PWCs without any CDM-related clinical signs, to determine the current allele frequency in Japan. Both of the assays provided clear-cut genotyping.…”