Digital PCR detection of plasmid DNA administered to the skeletal muscle of a microminipig: a model case study for gene doping detection

Digital PCR detection of plasmid DNA administered to the skeletal muscle of a microminipig : a model case study for gene doping detection / Teruaki Tozaki, Shiori Gamo, Masaki Takasu, Mio Kikuchi, Hironaga Kakoi, Kei-Ichi Hirota, Kanichi Kusano, Shun-Ichi Nagata. - (BMC Res Notes 11 (2018) 10 October)

  • PMID: 30309394
  • PMCID: PMC6180624
  • DOI: 10.1186/s13104-018-3815-6


Abstract

Objective: Doping control is an important and indispensable aspect of fair horse racing; genetic doping has been recently included to this. In this study, we aimed to develop a detection method of gene doping. A plasmid cloned with human erythropoietin gene (p.hEPO, 250 μg/head) was intramuscularly injected into a microminipig. Subsequently, p.hEPO was extracted from 1 mL of plasma and detected by droplet digital polymerase chain reaction.

Results: The results confirmed that the maximum amount of plasmid was detected at 15 min after administration and the majority of the plasmid was degraded in the bloodstream within 1-2 days after administration. In contrast, low amounts of p.hEPO were detected at 2-3 weeks after administration. These results suggest that the proposed method to detect gene doping can help obtain information for experiments using horses.

 

Original document

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Science
Study
Date
10 October 2018
People
Gamo, Shiori
Hirota, Kei-Ichi
Kakoi, Hironaga
Kikuchi, Mio
Kusano, Kanichi
Nagata, Shun-Ichi
Takasu, Masaki
Tozaki, Teruaki
Country
Japan
Language
English
Other organisations
Gifu University
Japan Racing Association (JRA)
Laboratory of Racing Chemistry (LRC)
Analytical aspects
Testing method development
Doping classes
M3. Gene And Cell Doping
Medical terms
Gene Therapy
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Scientific article
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Date generated
14 January 2021
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15 January 2021
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