Individual identification of racehorses from urine samples using a 26-plex single-nucleotide polymorphism assay.
Abstract: 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.
© 2012 American Academy of Forensic Sciences.
Publication Date: 2012-10-12 PubMed ID: 23061924DOI: 10.1111/j.1556-4029.2012.02291.xGoogle Scholar: Lookup
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- Journal Article
Summary
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The research article describes a new method for horse identification using a genotyping assay based on 26 single-nucleotide polymorphisms present in horse urine samples. This method seeks to improve the accuracy of postracing doping tests.
Objective of the Research
- The main goal of this research was to develop a system that can accurately determine the identity of individual horses from their urine samples. This system could be used to improve the efficiency and reliability of postracing doping tests.
Methodology
- The researchers used a 26-plex single-nucleotide polymorphism (SNP) assay for the genotyping. SNPs are variations at single points in the DNA sequence and are typically highly unique, making them useful identifiers.
- DNA was extracted from the urine samples using a commercially available DNA/RNA extraction kit.
- The genotyping was performed using a technique known as a SNaPshot, which can accurately identify SNPs to generate individual profiles.
- The researchers compared DNA profiles generated from the urine samples with profiles from blood and hair samples to verify their accuracy.
Outcomes
- The researchers found that the SNP assay was able to accurately identify individual horses within the Thoroughbred population. The probability of identity was extremely low, suggesting a high level of accuracy.
- Compared to conventional methods of horse identification, such as short tandem repeat assays, the SNP assay required less DNA and was successful in genotyping in 97% of cases.
- The researchers also found that the urinary DNA could still be properly genotyped under different storage conditions, such as refrigeration or repeated freezing and thawing.
Significance of the Study
- The results of this research suggest that the SNP assay can be a valuable tool for identifying individual horses from urine samples following races.
- As such, when suspicious results are obtained from horse doping tests, this system can be used to verify the identity of the tested horse, improving the integrity of the racing sport.
Cite This Article
APA
Kakoi H, Kijima-Suda I, Gawahara H, Kinoshita K, Tozaki T, Hirota K, Yoshizawa M.
(2012).
Individual identification of racehorses from urine samples using a 26-plex single-nucleotide polymorphism assay.
J Forensic Sci, 58(1), 21-28.
https://doi.org/10.1111/j.1556-4029.2012.02291.x Publication
Researcher Affiliations
- Genetic Analysis Section, Laboratory of Racing Chemistry, 1731-2, Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan. h-kakoi@lrc.or.jp
MeSH Terms
- Animals
- DNA / urine
- DNA Fingerprinting / methods
- DNA Fingerprinting / veterinary
- Genotype
- Horses / genetics
- Microsatellite Repeats
- Polymerase Chain Reaction
- Polymorphism, Single Nucleotide
Citations
This article has been cited 2 times.- Brooks A, Creighton EK, Gandolfi B, Khan R, Grahn RA, Lyons LA. SNP Miniplexes for Individual Identification of Random-Bred Domestic Cats.. J Forensic Sci 2016 May;61(3):594-606.
- Chen JW, Uboh CE, Soma LR, You Y, Jiang Z, Li X, Guan F, Liu Y. Identification of sample donor by 24-plex short tandem repeat in a post-race equine plasma containing dexamethasone.. Springerplus 2014;3:94.
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