Next Article in Journal
A Gold Standard, CRISPR/Cas9-Based Complementation Strategy Reliant on 24 Nucleotide Bookmark Sequences
Previous Article in Journal
Exploring the Mechanisms of Multiple Insecticide Resistance in a Highly Plasmodium-Infected Malaria Vector Anopheles funestus Sensu Stricto from Sahel of Northern Nigeria
Open AccessArticle

Microfluidic Quantitative PCR Detection of 12 Transgenes from Horse Plasma for Gene Doping Control

1
Genetic Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya, Tochigi 320-0851, Japan
2
Equine Department, Japan Racing Association, 6-11-1 Roppongi, Minato, Tokyo 106-8401, Japan
*
Author to whom correspondence should be addressed.
Both authors contributed equally to this manuscript.
Genes 2020, 11(4), 457; https://doi.org/10.3390/genes11040457
Received: 30 March 2020 / Revised: 13 April 2020 / Accepted: 20 April 2020 / Published: 23 April 2020
(This article belongs to the Section Animal Genetics and Genomics)
Gene doping, an activity which abuses and misuses gene therapy, is a major concern in sports and horseracing industries. Effective methods capable of detecting and monitoring gene doping are urgently needed. Although several PCR-based methods that detect transgenes have been developed, many of them focus only on a single transgene. However, numerous genes associated with athletic ability may be potential gene-doping material. Here, we developed a detection method that targets multiple transgenes. We targeted 12 genes that may be associated with athletic performance and designed two TaqMan probe/primer sets for each one. A panel of 24 assays was prepared and detected via a microfluidic quantitative PCR (MFQPCR) system using integrated fluidic circuits (IFCs). The limit of detection of the panel was 6.25 copy/μL. Amplification-specificity was validated using several concentrations of reference materials and animal genomic DNA, leading to specific detection. In addition, target-specific detection was successfully achieved in a horse administered 20 mg of the EPO transgene via MFQPCR. Therefore, MFQPCR may be considered a suitable method for multiple-target detection in gene-doping control. To our knowledge, this is the first application of microfluidic qPCR (MFQPCR) for gene-doping control in horseracing. View Full-Text
Keywords: gene doping; horse; microfluidic qPCR; multiple-target detection; transgene gene doping; horse; microfluidic qPCR; multiple-target detection; transgene
Show Figures

Figure 1

MDPI and ACS Style

Tozaki, T.; Ohnuma, A.; Kikuchi, M.; Ishige, T.; Kakoi, H.; Hirota, K.-I.; Kusano, K.; Nagata, S.-I. Microfluidic Quantitative PCR Detection of 12 Transgenes from Horse Plasma for Gene Doping Control. Genes 2020, 11, 457.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop