Detection of adeno-associated viral DNA in equine post-administration frozen blood and plasma samples after long-term storage.
Abstract: Gene doping in horses is a threat to the fairness in sport and has serious implications for animal welfare. To investigate the effect of long-term storage on the detection of AAV in plasma and whole blood, samples from an administration study using an adeno-associated virus serotype 6 expressing green fluorescence protein (AAV6-GFP) were stored at -20°C for 8 months before analysis. The AAV vector was detected in stored plasma samples, following the same detection profile as the fresh plasma samples. The stored blood showed lower overall DNA detection but followed the same detection profile as the plasma samples. This study provides confidence that re-analysing plasma samples and/or analysing a frozen 'B' sample with different matrix such as whole blood after prolonged storage will still result in the detection of gene doping material.
© 2023 LGC Ltd and British Horseracing Authority. Drug Testing and Analysis published by John Wiley & Sons Ltd.
Publication Date: 2023-09-06 PubMed ID: 37671588DOI: 10.1002/dta.3569Google Scholar: Lookup
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- Journal Article
Summary
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This research article explores the long-term detection of gene doping material in horse blood and plasma samples, providing confidence that regardless of storage duration, detection may still be possible, thereby maintaining fairness in sport and promoting animal wellbeing.
Overview of the Study
- The study’s primary aim is to investigate the effect of long-term storage on the detectability of AAVs (Adeno-associated viruses) in horse plasma and blood samples. This is driven by the concerns and implications of gene doping in sports, with specific emphasis on horse racing.
- The study used samples from an administration study involving an AAV serotype 6 expressing green fluorescence protein (AAV6-GFP) and stored these samples at -20°C for 8 months before proceeding with analysis.
Findings of the Study
- The research discovered that despite the long-term storage of plasma samples, the AAV vector was still detectable, matching the detection profile of fresh plasma samples.
- Although the stored blood showed a slightly lower DNA detection rate when compared to the plasma samples, the detection patterns across both samples remained consistent.
Implications of the Study
- The results help boost confidence regarding the re-analysis of plasma samples or analysis of frozen ‘B’ samples with a different matrix such as whole blood after prolonged storage. It provides assurance that the detection of gene doping material will still be successful, regardless of the storage duration.
- This research has significant implications in regulating sports like horse racing, as it can help in identifying and curtailing unfair practices such as gene doping. Moreover, it also has potentials to contribute to animal welfare by discouraging harmful doping practices.
Cite This Article
APA
Maniego J, Pesko B, Habershon-Butcher J, Hincks P, Taylor P, Stewart G, Proudman C, Ryder E.
(2023).
Detection of adeno-associated viral DNA in equine post-administration frozen blood and plasma samples after long-term storage.
Drug Test Anal.
https://doi.org/10.1002/dta.3569 Publication
Researcher Affiliations
- Sport and Specialised Analytical Services, LGC, Fordham, UK.
- Sport and Specialised Analytical Services, LGC, Fordham, UK.
- British Horseracing Authority, London, UK.
- Sport and Specialised Analytical Services, LGC, Fordham, UK.
- Sport and Specialised Analytical Services, LGC, Fordham, UK.
- School of Biosciences and Medicine, University of Surrey, Guildford, UK.
- School of Veterinary Medicine, University of Surrey, Guildford, UK.
- Sport and Specialised Analytical Services, LGC, Fordham, UK.
Grant Funding
- British Horseracing Authority (BHA)
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This article includes 15 references
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