Detection of Gene Doping Using Dried Blood Spots from a Mouse Model with rAAV9 Vector-Mediated Human Erythropoietin Expression as a Pilot Study

Analytica Pub Date : 2024-06-04 DOI:10.3390/analytica5020017
Norihiro Otani, Yasuharu Kanki, Kieu D. M. Nguyen, T. Sugasawa
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Abstract

Rapid advancements in gene technology have raised concerns regarding the potential abuse of techniques, such as gene doping, for enhancing athletic performance. To identify this possibility, a reliable procedure for detecting doping genes is required. Although detection methods for doping genes have been created, there are still areas for further improvement. One significant challenge is the high storage and transport costs of the test samples. For this issue, the dried blood spot (DBS) method can be a cost-effective solution. This study aimed to assess the practicality of incorporating DBSs into the gene doping detection process as a pilot study. Whole-blood samples were initially collected from mice engineered to express human erythropoietin from the rAAV vector. Then, the blood was placed in filter papers and left to dry at room temperature for five hours to form DBSs. These DBSs were subsequently preserved in sealed plastic bags at room temperature. After the extraction of DNA, DBSs were formed, and TaqMan-qPCR was utilized to detect the presence of rAAV vector-derived DNA. The finding confirmed that doping gene-specific fragments were successfully detected in DBSs. This outcome suggests that the DBS method is an effective approach to be considered when developing a comprehensive protocol for gene doping detection.
利用 rAAV9 载体介导的人类促红细胞生成素表达小鼠模型的干血斑检测基因兴奋剂的先导研究
基因技术的飞速发展引发了人们对可能滥用基因兴奋剂等技术提高运动成绩的担忧。为了识别这种可能性,需要一种可靠的程序来检测兴奋剂基因。虽然兴奋剂基因的检测方法已经问世,但仍有需要进一步改进的地方。其中一个重大挑战是检测样本的储存和运输成本较高。针对这一问题,干血斑(DBS)法不失为一种经济有效的解决方案。本研究旨在评估将 DBS 纳入基因兴奋剂检测流程的实用性。首先从利用 rAAV 载体表达人促红细胞生成素的小鼠身上采集全血样本。然后,将血液置于滤纸中,在室温下晾干五小时,形成 DBSs。这些 DBS 随后被密封在室温下的塑料袋中保存。提取 DNA 后,形成 DBS,并利用 TaqMan-qPCR 检测 rAAV 载体 DNA 的存在。结果证实,在 DBSs 中成功检测到了掺杂基因特异性片段。这一结果表明,在制定全面的基因掺杂检测方案时,DBS 方法是一种值得考虑的有效方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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CiteScore
1.80
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