With the rapid progress of genetic engineering and gene therapy, theWorld Anti-Doping\nAgency has been alerted to gene doping and prohibited its use in sports. However, there is no\nstandard method available yet for the detection of transgenes delivered by recombinant adenoviral\n(rAdV) vectors. Here, we aim to develop a detection method for transgenes delivered by rAdV\nvectors in a mouse model that mimics gene doping. These rAdV vectors containing the mCherry gene\nwas delivered in mice through intravenous injection or local muscular injection. After five days, stool\nand whole blood samples were collected, and total DNA was extracted. As additional experiments,\nwhole blood was also collected from the mouse tail tip until 15 days from injection of the rAdv\nvector. Transgene fragments from different DNA samples were analyzed using semi-quantitative\nPCR (sqPCR), quantitative PCR (qPCR), and droplet digital PCR (ddPCR). In the results, transgene\nfragments could be directly detected from blood cell fraction DNA, plasma cell-free DNA, and stool\nDNA by qPCR and ddPCR, depending on specimen type and injection methods. We observed that a\ncombination of blood cell fraction DNA and ddPCR was more sensitive than other combinations used\nin this model. These results could accelerate the development of detection methods for gene doping.
Loading....