Individual identification of racehorses from urine samples using a 26-plex single-nucleotide polymorphism assay.
To construct a system for identifying individual horses from urine samples that are submitted for postracing doping tests, we developed a genotyping assay based on 26-plex single-nucleotide polymorphisms (SNPs). DNA was isolated from urine using a commercially available DNA/RNA extraction kit, and SNP genotyping was achieved with a SNaPshot(™) technique. DNA profiles including 26 SNPs were acquired from urine samples and blood/hair samples. Within the studied Thoroughbred population, the 26-plex assay showed a probability of identity of 5.80 × 10(-11). Compared to the conventional short tandem repeat assay, the SNP assay used less DNA, and the rate of successful genotyping was improved to 97% using aliquots of horse urine as small as 140 μL. The urinary DNA could be successfully genotyped under proper storage concerning refrigeration or freeze-thawing. This SNP assay can be used for individual identification when suspicious results are obtained from horse doping tests.
Genetic Analysis Section, Laboratory of Racing Chemistry, 1731-2, Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan. h-kakoi@lrc.or.jp
Hironaga
H
Kakoi
Isao
I
Kijima-Suda
Hitoshi
H
Gawahara
Kenji
K
Kinoshita
Teruaki
T
Tozaki
Kei-ichi
K
Hirota
Midori
M
Yoshizawa
D000818:Animals; D004247:DNA / Q000652:urine*; D016172:DNA Fingerprinting / Q000379:methods / Q000662:veterinary*; D005838:Genotype; D006736:Horses / Q000235:genetics*; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide
D004247:DNA