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Journal of Veterinary Diagnostic Investigation, Vol 16, Issue 4, 299-304
Copyright © 2004 by American Association of Veterinary Laboratory Diagnosticians

Evaluation Studies

Comparison of polymerase chain reaction-restriction fragment length polymorphism assay and enzyme assay for diagnosis of G(M1)-gangliosidosis in Shiba dogs

O Yamato, A Kobayashi, H Satoh, D Endoh, T Shoda, Y Masuoka, A Hatakeyama, EO Jo, T Asano, M Yonemura, M Yamasaki, and Y Maede

Laboratory of Internal Medicine, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

In the present study, diagnostic methods for canine G(M1)-gangliosidosis were examined by comparing a DNA mutation assay with an enzyme assay. Sixty-two Shiba dogs of a pedigree with G(M1)-gangliosidosis were differentiated into 3 genotypes, i.e., normal, heterozygous, and homozygous affected dogs, using a DNA mutation assay, which consists of polymerase chain reaction amplification and the determination of restriction fragment length polymorphisms. The beta-galactosidase activity in leukocytes, umbilical cords, and plasma was measured using 4-methylumbelliferyl beta-D-galactoside and p-nitrophenyl beta-D-galactoside as artificial substrates and compared among the 3 genotypes. The results showed that it was possible to identify homozygous dogs with the enzyme assay using leukocytes and umbilical cords. When using leukocytes, heterozygous carriers could be differentiated from normal dogs in many cases. However, the use of the DNA mutation assay is essential for a complete determination of heterozygous carriers because of the overlap in the distribution of enzyme activity between these 2 groups. When umbilical cords were used, heterozygous carriers could not be differentiated from normal dogs because of no significant difference in enzyme activity between these 2 groups. The beta-galactosidase activity in plasma was not applicable to the diagnosis and genotyping of G(M1)-gangliosidosis in Shiba dogs.


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O. Yamato, E.-O. Jo, H.-S. Chang, H. Satoh, T. Shoda, R. Sato, M. Uechi, N. Kawasaki, Y. Naito, M. Yamasaki, et al.
Molecular screening of canine GM1 gangliosidosis using blood smear specimens after prolonged storage: detection of carriers among shiba dogs in northern Japan
J Vet Diagn Invest, January 1, 2008; 20(1): 68 - 71.
[Abstract] [Full Text] [PDF]


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