Cascella R, Strafella C, Ragazzo M, Zampatti S, Borgiani P, Gambardella S, Pirazzoli A, Novelli G, Giardina E
Department of Biomedicine and Prevention, School of Medicine, University of Rome 'Tor Vergata', Rome, Italy.
Neuromed IRCCS, Pozzilli, Italy.
Pharmacogenomics J. 2015 Apr;15(2):196-200. doi: 10.1038/tpj.2014.48. Epub 2014 Sep 9.
One of the most successful applications of pharmacogenetics research is the genetic screening for HLA-B57:01, strongly associated with an increased risk to develop hypersensitivity reaction in HIV-positive patients following abacavir administration. Taking into consideration the limits of current genotyping methodologies, we have developed and validated (150 buccal swabs) an inexpensive pharmacogenetic approach for HLA-B57:01 typing. In our assay DNA extraction and amplification are combined in one single step (direct PCR protocol), which is performed directly on the biological sample without the need of extraction and sequencing passages. The amplicons obtained by direct PCR can be easily separated on the agarose gel under ultraviolet. As per our results, the direct PCR represents a good alternative to the traditional methods of HLA-B*57:01 pharmacogenetic test, especially for those laboratories or countries where currently available approaches are often not available or not affordable. Furthermore it is an innovative approach, promoting a personalized, safer and cost-effective therapy.
药物遗传学研究最成功的应用之一是对HLA - B57:01进行基因筛查，该基因与HIV阳性患者服用阿巴卡韦后发生超敏反应风险增加密切相关。考虑到当前基因分型方法的局限性，我们开发并验证了（150份口腔拭子样本）一种用于HLA - B57:01分型的低成本药物遗传学方法。在我们的检测方法中，DNA提取和扩增合并为一个步骤（直接PCR方案），直接在生物样本上进行，无需提取和测序步骤。通过直接PCR获得的扩增子在紫外线下可轻松在琼脂糖凝胶上分离。根据我们的结果，直接PCR是HLA - B*57:01药物遗传学检测传统方法的良好替代方法，尤其适用于那些目前常用方法不可用或负担不起的实验室或国家。此外，它是一种创新方法，有助于推动个性化、更安全且具成本效益的治疗。
