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.
