Digital PCR detection of plasmid DNA administered to the skeletal muscle of a microminipig: a model case study for gene doping detection
2018
Teruaki Tozaki | Shiori Gamo | Masaki Takasu | Mio Kikuchi | Hironaga Kakoi | Kei-ichi Hirota | Kanichi Kusano | Shun-ichi Nagata
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.
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