Development of multiplexed screening and confirmatory assays for gene doping transgenes in human plasma within an ISO17025 accredited laboratory

Code: 253E03ER

Gene doping is a growing concern in athletic sports for both fair play and potential adverse health implications. One likely method of gene doping involves gene transfer: the introduction of additional copies of performance-related genes into cells by injection of a carrier virus. The method of packing of these genes into the virus creates unique sites that are not present in the human genome. These unique sites form the basis of the gene doping detection assays. By harnessing the parallel power of targeted next-generation sequencing (NGS), we can combine a suite of detection assays into one high-throughput screening test, greatly reducing costs compared to single target methods.

This project will construct a pool of 10 gene doping candidate target assays and test them on human plasma samples using a pipeline that has already been accredited to ISO 17025 for equine samples. For confirmatory analysis, single-target qPCR assays will be employed in accordance with current WADA guidelines on technology use. In addition, qPCR products are sequenced to verify using our novel method.

As a ‘proof of principle’ screen, human plasma samples available from commercial suppliers will be processed and assessed alongside spiked samples. For each gene, a confirmatory qPCR assay will be designed and tested, culminating in mock confirmatory tests as a proof of principle for future regulated validation.