論文

査読有り 国際誌
2019年3月21日

Droplet Digital PCR Detection of the Erythropoietin Transgene from Horse Plasma and Urine for Gene-Doping Control.

Genes
  • Teruaki Tozaki
  • ,
  • Aoi Ohnuma
  • ,
  • Masaki Takasu
  • ,
  • Mio Kikuchi
  • ,
  • Hironaga Kakoi
  • ,
  • Kei-Ichi Hirota
  • ,
  • Kanichi Kusano
  • ,
  • Shun-Ichi Nagata

10
3
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.3390/genes10030243

Indiscriminate genetic manipulation to improve athletic ability is a major threat to human sports and the horseracing industry, in which methods involving gene-doping, such as transgenesis, should be prohibited to ensure fairness. Therefore, development of methods to detect indiscriminate genetic manipulation are urgently needed. Here, we developed a highly sensitive method to detect horse erythropoietin (EPO) transgenes using droplet digital PCR (ddPCR). We designed two TaqMan probe/primer sets, and the EPO transgene was cloned into a plasmid for use as a model. We extracted the spiked EPO transgene from horse plasma and urine via magnetic beads, followed by ddPCR amplification for absolute quantification and transgene detection. The results indicated high recovery rates (at least ~60% and ~40% in plasma and urine, respectively), suggesting successful detection of the spiked transgene at concentrations of >130 and 200 copies/mL of plasma and urine, respectively. Additionally, successful detection was achieved following intramuscular injection of 20 mg of the EPO transgene. This represents the first study demonstrating a method for detecting the EPO transgene in horse plasma and urine, with our results demonstrating its efficacy for promoting the control of gene-doping in the horseracing industry.

リンク情報
DOI
https://doi.org/10.3390/genes10030243
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/30901981
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6471249
ID情報
  • DOI : 10.3390/genes10030243
  • PubMed ID : 30901981
  • PubMed Central 記事ID : PMC6471249

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