Digital PCR Detection of Plasmid DNA Administered to the Skeletal Muscle of a Microminipig: a Model Case Study for Gene Doping Detection

Overview

Journal BMC Res Notes

Publisher Biomed Central

Specialties Biology
General Medicine

Date 2018 Oct 13

PMID 30309394

Citations 13

Authors

Teruaki Tozaki

Shiori Gamo

Masaki Takasu

Mio Kikuchi

Hironaga Kakoi

Kei-Ichi Hirota

Kanichi Kusano

Shun-Ichi Nagata

Affiliations

Soon will be listed here.

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.

Citing Articles

Gene doping in horse racing and equine sports: Current landscape and future perspectives.

Puchalska M, Witkowska-Pilaszewicz O Equine Vet J. 2024; 57(2):312-324.

PMID: 39267222 PMC: 11807943. DOI: 10.1111/evj.14418.

Administration and detection of a multi-target rAAV gene doping vector in horses using multiple matrices and molecular techniques.

Maniego J, Harding C, Habershon-Butcher J, Hincks P, Ryder E Gene Ther. 2024; 31(9-10):477-488.

PMID: 38972888 DOI: 10.1038/s41434-024-00462-0.

Detection of Indiscriminate Genetic Manipulation in Thoroughbred Racehorses by Targeted Resequencing for Gene-Doping Control.

Tozaki T, Ohnuma A, Nakamura K, Hano K, Takasu M, Takahashi Y Genes (Basel). 2022; 13(9).

PMID: 36140757 PMC: 9498419. DOI: 10.3390/genes13091589.

Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector.

Sugasawa T, Nakano T, Fujita S, Matsumoto Y, Ishihara G, Aoki K Genes (Basel). 2021; 12(8).

PMID: 34440425 PMC: 8392868. DOI: 10.3390/genes12081249.

Rare and common variant discovery by whole-genome sequencing of 101 Thoroughbred racehorses.

Tozaki T, Ohnuma A, Kikuchi M, Ishige T, Kakoi H, Hirota K Sci Rep. 2021; 11(1):16057.

PMID: 34362995 PMC: 8346562. DOI: 10.1038/s41598-021-95669-1.

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