Individual Identification of Racehorses from Urine Samples Using a 26-Plex Single-Nucleotide Polymorphism Assay

被引:3
|
作者
Kakoi, Hironaga [1 ]
Kijima-Suda, Isao [2 ]
Gawahara, Hitoshi [1 ]
Kinoshita, Kenji [2 ]
Tozaki, Teruaki [3 ]
Hirota, Kei-ichi [3 ]
Yoshizawa, Midori [4 ]
机构
[1] Lab Racing Chem, Genet Anal Sect, Utsunomiya, Tochigi 3200851, Japan
[2] Lab Racing Chem, Doping Anal Sect, Utsunomiya, Tochigi 3200851, Japan
[3] Lab Racing Chem, Mol Genet Sect, Utsunomiya, Tochigi 3200851, Japan
[4] Utsunomiya Univ, Dept Anim Sci, Fac Agr, Utsunomiya, Tochigi 3218505, Japan
关键词
forensic science; DNA typing; horse; urine; single-nucleotide polymorphism; individual identification; PARENTAGE EXCLUSION; THOROUGHBRED HORSES; MICROSATELLITE DNA; MARKERS; VALIDATION; LOCI; TECHNOLOGIES; PROBABILITY; KIT;
D O I
10.1111/j.1556-4029.2012.02291.x
中图分类号
DF [法律]; D9 [法律]; R [医药、卫生];
学科分类号
0301 ; 10 ;
摘要
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 (TM) 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 x 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 mu L. The urinary DNA could be successfully genotyped under proper storage concerning refrigeration or freezethawing. This SNP assay can be used for individual identification when suspicious results are obtained from horse doping tests.
引用
收藏
页码:21 / 28
页数:8
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