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

BMC Res Notes. 2018 Oct 10;11(1):708. 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.

Keywords: Gene doping; Horseracing; Plasmid; Thoroughbred.

MeSH terms

  • Animals
  • DNA Fragmentation
  • Doping in Sports / methods
  • Doping in Sports / prevention & control*
  • Erythropoietin / blood
  • Erythropoietin / genetics*
  • Erythropoietin / pharmacokinetics
  • Gene Expression
  • Horses
  • Humans
  • Injections, Intramuscular
  • Male
  • Plasmids / administration & dosage
  • Plasmids / chemistry*
  • Plasmids / metabolism
  • Polymerase Chain Reaction / methods*
  • Recombinant Proteins / blood
  • Recombinant Proteins / genetics
  • Recombinant Proteins / pharmacokinetics
  • Sensitivity and Specificity
  • Swine
  • Swine, Miniature
  • Transgenes

Substances

  • EPO protein, human
  • Recombinant Proteins
  • Erythropoietin