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

Abstract:

: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.

journal_name

Genes (Basel)

journal_title

Genes

authors

Tozaki T,Ohnuma A,Kikuchi M,Ishige T,Kakoi H,Hirota KI,Kusano K,Nagata SI

doi

10.3390/genes11040457

subject

Has Abstract

pub_date

2020-04-23 00:00:00

issue

4

issn

2073-4425

pii

genes11040457

journal_volume

11

pub_type

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