The identification of polymorphic microsatellite loci in the horse and their use in thoroughbred parentage testing.
Abstract: Six new horse microsatellite loci were identified by sequencing M13 clones containing horse genomic inserts which gave positive signals when probed with a CA/GT repeat probe. Oligonucleotide primer pairs were synthesized for these loci and for two previously described horse microsatellites, HTG4 and HTG6. Polymerase chain reaction assays were then carried out on a panel of 20 different unrelated Thoroughbred horse DNAs. DNAs from eight cases of double covering which could not be solved by conventional blood typing were also examined. Several of the loci amplified were found to be polymorphic and, using a limited subset of primers, a clear exclusion could be established for one of the stallions in five of the cases. DNA typing is therefore a useful adjunct to blood typing in the horse and indeed, in the future will probably replace it.
Publication Date: 1995-01-01 PubMed ID: 7735875DOI: 10.1016/s0007-1935(05)80057-0Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research identified six new horse genetic markers (microsatellites) that can clearly and accurately determine the parents of thoroughbred horses, providing a more accurate approach than the existing blood typing methods.
Introduction and Methodology
- The research involved sequencing M13 clones that contained horse genomic inserts. When these inserts were probed with a specific type of marker (CA/GT repeat probe), they produced positive signals, identifying them as potential microsatellite loci (positions within the genetic code).
- Scientists then synthesized “oligonucleotide primer pairs” for these loci, along with two already known horse microsatellites, HTG4 and HTG6. These primers enable the next step, which is polymerase chain reaction (PCR) assay.
- PCR assays were carried out on DNA from 20 unrelated Thoroughbred horses. This process amplified the regions of DNA defined by the primer pairs, helping researchers confirm these regions as new microsatellite loci.
Testing on Cases of Double Covering
- The researchers further examined DNA samples from eight instances of ‘double covering’, a situation where a mare may have mated with two different stallions within a short period.
- The conventional method of blood typing was unable to definitively allocate parentage in these instances.
- By using the identified microsatellite markers, the team could establish a clear parental lineage in five of those eight cases.
Conclusion and Implications
- Several of the amplified loci were polymorphic, which means they had multiple forms, making them suitable for identifying genetic variation and thus, verifying lineage.
- The study proved that DNA typing using microsatellite markers could provide a reliable method to solve complex paternity cases in horse breeding. This method was found to be a useful supplement to the current blood typing method.
- The researchers suggest that DNA typing could likely replace blood typing in the future, offering a more accurate and reliable system for determining horse lineage and parentage.
Cite This Article
APA
Binns MM, Holmes NG, Holliman A, Scott AM.
(1995).
The identification of polymorphic microsatellite loci in the horse and their use in thoroughbred parentage testing.
Br Vet J, 151(1), 9-15.
https://doi.org/10.1016/s0007-1935(05)80057-0 Publication
Researcher Affiliations
- Animal Health Trust, Kennett, Newmarket, Suffolk, UK.
MeSH Terms
- Animals
- Base Sequence
- Breeding
- DNA Primers
- DNA, Satellite / genetics
- Horses / genetics
- Molecular Sequence Data
- Pedigree
- Polymerase Chain Reaction
- Polymorphism, Genetic / genetics
Citations
This article has been cited 16 times.- Yordanov G, Mehandjyiski I, Palova N, Atsenova N, Neov B, Radoslavov G, Hristov P. Genetic Diversity and Structure of the Main Danubian Horse Paternal Genealogical Lineages Based on Microsatellite Genotyping. Vet Sci 2022 Jul 1;9(7).
- de Groot M, Anderson H, Bauer H, Bauguil C, Bellone RR, Brugidou R, Buckley RM, Dovč P, Forman O, Grahn RA, Kock L, Longeri M, Mouysset-Geniez S, Qiu J, Sofronidis G, van der Goor LHP, Lyons LA. Standardization of a SNP panel for parentage verification and identification in the domestic cat (Felis silvestris catus). Anim Genet 2021 Oct;52(5):675-682.
- Martinez MM, Costa M, Ratti C. Molecular screening of XY SRY-negative sex reversal cases in horses revealed anomalies in amelogenin testing. J Vet Diagn Invest 2020 Nov;32(6):938-941.
- Yatkın S, Özdil F, Ünal EÖ, Genç S, Kaplan S, Gürcan EK, Arat S, Soysal Mİ. Genetic Characterization of Native Donkey (Equus asinus) Populations of Turkey Using Microsatellite Markers. Animals (Basel) 2020 Jun 24;10(6).
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- Lee SY, Cho GJ. Parentage testing of Thoroughbred horse in Korea using microsatellite DNA typing. J Vet Sci 2006 Mar;7(1):63-7.
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- Jlassi M, Dhifalli I, Ouled Ahmed H, Lasfar F, El Gtari M, Jemmali B. Genetic Characterization of the Arabian Horse Population in Tunisia Using Microsatellites. Life (Basel) 2025 Dec 16;15(12).
- Kim D, Lee S, Oyungerel B, Cho G. Evaluation of the Effectiveness of Single-Nucleotide Polymorphisms Versus Microsatellites for Parentage Verification in Horse Breeds. Vet Sci 2025 Sep 15;12(9).
- Yordanov G, Yordanov T, Mehandjyiski I, Radoslavov G, Salkova D, Hristov P. Population Structure and Genetic Diversity Among Shagya Arabian Horse Genealogical Lineages in Bulgaria Based on Microsatellite Genotyping. Vet Sci 2025 Aug 19;12(8).
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