Real-time PCR genotyping assay for canine progressive rod-cone degeneration and mutant allele frequency in Toy Poodles, Chihuahuas and Miniature Dachshunds in Japan

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  • KOHYAMA Moeko
    Laboratory of Clinical Pathology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1–21–24 Kohrimoto, Kagoshima 890–0065, Japan
  • TADA Naomi
    International Institute of Small Animal Medicine (Bio Plus), AHB Inc., 3–7–11 Kiba, Koutou-ku, Tokyo 135–0042, Japan
  • MITSUI Hiroko
    International Institute of Small Animal Medicine (Bio Plus), AHB Inc., 3–7–11 Kiba, Koutou-ku, Tokyo 135–0042, Japan
  • TOMIOKA Hitomi
    International Institute of Small Animal Medicine (Bio Plus), AHB Inc., 3–7–11 Kiba, Koutou-ku, Tokyo 135–0042, Japan
  • TSUTSUI Toshihiko
    International Institute of Small Animal Medicine (Bio Plus), AHB Inc., 3–7–11 Kiba, Koutou-ku, Tokyo 135–0042, Japan
  • YABUKI Akira
    Laboratory of Clinical Pathology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1–21–24 Kohrimoto, Kagoshima 890–0065, Japan
  • RAHMAN Mohammad Mahbubur
    Laboratory of Clinical Pathology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1–21–24 Kohrimoto, Kagoshima 890–0065, Japan
  • KUSHIDA Kazuya
    Laboratory of Clinical Pathology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1–21–24 Kohrimoto, Kagoshima 890–0065, Japan
  • MIZUKAMI Keijiro
    Laboratory of Clinical Pathology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1–21–24 Kohrimoto, Kagoshima 890–0065, Japan
  • YAMATO Osamu
    Laboratory of Clinical Pathology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1–21–24 Kohrimoto, Kagoshima 890–0065, Japan

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Abstract

Canine progressive rod-cone degeneration (PRCD) is a middle- to late-onset, autosomal recessive, inherited retinal disorder caused by a substitution (c.5G>A) in the canine PRCD gene that has been identified in 29 or more purebred dogs. In the present study, a TaqMan probe-based real-time PCR assay was developed and evaluated for rapid genotyping and large-scale screening of the mutation. Furthermore, a genotyping survey was carried out in a population of the three most popular breeds in Japan (Toy Poodles, Chihuahuas and Miniature Dachshunds) to determine the current mutant allele frequency. The assay separated all the genotypes of canine PRCD rapidly, indicating its suitability for large-scale surveys. The results of the survey showed that the mutant allele frequency in Toy Poodles was high enough (approximately 0.09) to allow the establishment of measures for the prevention and control of this disorder in breeding kennels. The mutant allele was detected in Chihuahuas for the first time, but the frequency was lower (approximately 0.02) than that in Toy Poodles. The mutant allele was not detected in Miniature Dachshunds. This assay will allow the selective breeding of dogs from the two most popular breeds (Toy Poodle and Chihuahua) in Japan and effective prevention or control of the disorder.

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