建立一种用于直接检测基因兴奋剂的新型单拷贝引物-内含子跨越PCR（spiPCR）方法。

Establishing a novel single-copy primer-internal intron-spanning PCR (spiPCR) procedure for the direct detection of gene doping.

作者信息

Beiter Thomas, Zimmermann Martina, Fragasso Annunziata, Armeanu Sorin, Lauer Ulrich M, Bitzer Michael, Su Hua, Young William L, Niess Andreas M, Simon Perikles

机构信息

Department of Sports Medicine, Medical Clinic, University of Tüebingen, Germany.

出版信息

Exerc Immunol Rev. 2008;14:73-85.

PMID:19203085

Abstract

So far, the abuse of gene transfer technology in sport, so-called gene doping, is undetectable. However, recent studies in somatic gene therapy indicate that long-term presence of transgenic DNA (tDNA) following various gene transfer protocols can be found in DNA isolated from whole blood using conventional PCR protocols. Application of these protocols for the direct detection of gene doping would require almost complete knowledge about the sequence of the genetic information that has been transferred. Here, we develop and describe the novel single-copy primer-internal intron-spanning PCR (spiPCR) procedure that overcomes this difficulty. Apart from the interesting perspectives that this spiPCR procedure offers in the fight against gene doping, this technology could also be of interest in biodistribution and biosafety studies for gene therapeutic applications.

摘要

到目前为止，基因转移技术在体育领域的滥用，即所谓的基因兴奋剂，尚无法检测到。然而，近期体细胞基因治疗的研究表明，使用传统聚合酶链反应（PCR）方法，从全血中分离的DNA中可发现各种基因转移方案后转基因DNA（tDNA）的长期存在。将这些方法应用于直接检测基因兴奋剂需要几乎完全了解已转移的遗传信息序列。在此，我们开发并描述了一种新型的单拷贝引物-内含子跨越PCR（spiPCR）方法，该方法克服了这一困难。除了这种spiPCR方法在打击基因兴奋剂方面提供的有趣前景外，该技术在基因治疗应用的生物分布和生物安全性研究中也可能具有重要意义。