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基于 PCR 的基因转移载体检测:在基因兴奋剂检测中的应用。

PCR-based detection of gene transfer vectors: application to gene doping surveillance.

机构信息

Department of Molecular Genetics and Microbiology, University of Florida, College of Medicine, 1600 SW Archer Road, Gainesville, FL, 32610-0266, USA.

出版信息

Anal Bioanal Chem. 2013 Dec;405(30):9641-53. doi: 10.1007/s00216-013-7264-8. Epub 2013 Aug 4.

Abstract

Athletes who illicitly use drugs to enhance their athletic performance are at risk of being banned from sports competitions. Consequently, some athletes may seek new doping methods that they expect to be capable of circumventing detection. With advances in gene transfer vector design and therapeutic gene transfer, and demonstrations of safety and therapeutic benefit in humans, there is an increased probability of the pursuit of gene doping by athletes. In anticipation of the potential for gene doping, assays have been established to directly detect complementary DNA of genes that are top candidates for use in doping, as well as vector control elements. The development of molecular assays that are capable of exposing gene doping in sports can serve as a deterrent and may also identify athletes who have illicitly used gene transfer for performance enhancement. PCR-based methods to detect foreign DNA with high reliability, sensitivity, and specificity include TaqMan real-time PCR, nested PCR, and internal threshold control PCR.

摘要

运动员如果使用药物来提高运动表现,他们可能会有被禁止参加体育比赛的风险。因此,一些运动员可能会寻求新的兴奋剂方法,他们希望这些方法能够逃避检测。随着基因转移载体设计和治疗性基因转移的进步,以及在人类中展示的安全性和治疗益处,运动员进行基因兴奋剂的可能性增加了。为了预防基因兴奋剂的潜在风险,已经建立了检测方法,以直接检测用于兴奋剂的基因的 cDNA,以及载体控制元件。开发能够在运动中暴露基因兴奋剂的分子检测方法可以作为一种威慑手段,也可以识别出那些非法使用基因转移来提高成绩的运动员。用于检测具有高可靠性、灵敏度和特异性的外源 DNA 的基于 PCR 的方法包括 TaqMan 实时 PCR、嵌套 PCR 和内部阈值控制 PCR。

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