Identification of racehorse and sample contamination by novel 24-plex STR system.
Abstract: Proper identification of racehorses competing in an official race and maintenance of defensible chain of custody are important in doping control regulations. The purpose of this study was to develop a reliable multiplex PCR method for providing genetic evidence for matching donors to test samples by using short tandem repeat (STR) loci. Amplification of 21 STR loci from blood, urine or hair root was achieved in a single tube and STR length polymorphism was analyzed using fluorescent labeled capillary electrophoresis. This novel approach showed an allele confidence interval of 0.19-0.43 bp and size estimation error of 0-0.48 bp. In 90 thoroughbred (TB) and 171 standardbred (STB) horses, the method was highly discriminating and reproducible with probability of false identification of 1 in 10(11) (TB) and 1 in 10(13) (STB). All loci were highly polymorphic with an average probability of identity of 0.18 (TB) and 0.13 (STB), heterozygosity of 0.65 (TB) and 0.68 (STB), and polymorphism information content (PIC) of 0.62 (TB) and 0.69 (STB). The highest allele frequency also reflected the degree of polymorphism due to high correlation with PIC. To obtain evidence of sample tampering with human material, three human specific STR markers were included in the panel. This method is the first in the horseracing industry, specifically designed for racehorse identification and detection of equine sample contamination by human DNA.
Published by Elsevier Ireland Ltd.
Publication Date: 2009-09-02 PubMed ID: 20215027DOI: 10.1016/j.fsigen.2009.08.001Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article presents a new method for confirming the identity of racehorses and detecting any sample contamination. The technique uses a 24-plex system of short tandem repeat (STR) loci that can identify horses with a high degree of accuracy and detect if human DNA has contaminated a sample.
Overview of Methodology
- The focus of the study is to establish a dependable method of matching donated samples to their respective racehorses by amplifying 21 short tandem repeat (STR) loci from blood, urine, or hair root in a single-tube multiplex PCR method.
- The STR length polymorphism was analyzed using fluorescent labeled capillary electrophoresis which allows variability in the sequence of the DNA to be accurately gauged. This method utilizes the variations in repeated DNA sequences to create a unique genetic fingerprint for each individual.
- Three human-specific STR markers were also included in the 24-plex system to detect any human DNA that might have contaminated the horse samples.
Results of the Research
- The research indicates that the technique is highly accurate with an allele confidence interval ranging between 0.19-0.43 bp and a size estimation error of 0-0.48 bp.
- The method was tested on 90 thoroughbred (TB) and 171 standardbred (STB) horses, revealing a very low probability of false identification – 1 in 10(11) (TB) and 1 in 10(13) (STB).
- All the selected loci were highly polymorphic, showing a high level of variation, with the average probability of identity being 0.18 (TB) and 0.13 (STB), heterozygosity of 0.65 (TB) and 0.68 (STB), and polymorphism information content (PIC) of 0.62 (TB) and 0.69 (STB).
- The allele frequency also underscored the polymorphism degree as it showed a strong correlation with the PIC.
Significance and Utility of the Research
- This methodology is the first in the horse racing industry specifically developed for the identification of racehorses and the detection of equine sample contamination by human DNA.
- Reliable horse identification and detection of sample contamination are critical to enforce doping control regulations in racehorse events, making this research significant to ensure fairness in the sport.
Cite This Article
APA
Chen JW, Uboh CE, Soma LR, Li X, Guan F, You Y, Liu Y.
(2009).
Identification of racehorse and sample contamination by novel 24-plex STR system.
Forensic Sci Int Genet, 4(3), 158-167.
https://doi.org/10.1016/j.fsigen.2009.08.001 Publication
Researcher Affiliations
- University of Pennsylvania School of Veterinary Medicine, New Bolton Center Campus, Kennett Square, PA 19348, USA.
MeSH Terms
- Animals
- DNA Fingerprinting / veterinary
- Electrophoresis, Capillary
- Gene Frequency
- Genetic Markers
- Hair / metabolism
- Horses / genetics
- Humans
- Polymerase Chain Reaction
- Polymorphism, Genetic
- Sequence Analysis, DNA
- Tandem Repeat Sequences
Citations
This article has been cited 4 times.- Liu Y, Xu J, Chen M, Wang C, Li S. A unified STR profiling system across multiple species with whole genome sequencing data. BMC Bioinformatics 2019 Dec 20;20(Suppl 24):671.
- Arenas M, Pereira F, Oliveira M, Pinto N, Lopes AM, Gomes V, Carracedo A, Amorim A. Forensic genetics and genomics: Much more than just a human affair. PLoS Genet 2017 Sep;13(9):e1006960.
- Imtiaz A, Naz S. A Rapid and Cost-Effective Protocol for Screening Known Genes for Autosomal Recessive Deafness. Pak J Zool 2012 Jun 1;44(3):641-647.
- 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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