Lobato Ivan Magriñá, O'Sullivan Ciara K
INTERFIBIO Consolidated Research Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Països Catalans, 26, 43007, Tarragona, Spain.
Institució Catalana de Recerca i Estudis Avançats, Passeig Lluís Companys, 23, 08010 Barcelona, Spain.
Trends Analyt Chem. 2018 Jan;98:19-35. doi: 10.1016/j.trac.2017.10.015. Epub 2017 Oct 26.
Recombinase polymerase amplification (RPA) is a highly sensitive and selective isothermal amplification technique, operating at 37-42°C, with minimal sample preparation and capable of amplifying as low as 1-10 DNA target copies in less than 20 min. It has been used to amplify diverse targets, including RNA, miRNA, ssDNA and dsDNA from a wide variety of organisms and samples. An ever increasing number of publications detailing the use of RPA are appearing and amplification has been carried out in solution phase, solid phase as well as in a bridge amplification format. Furthermore, RPA has been successfully integrated with different detection strategies, from end-point lateral flow strips to real-time fluorescent detection amongst others. This review focuses on the different methodologies and advances related to RPA technology, as well as highlighting some of the advantages and drawbacks of the technique.
重组酶聚合酶扩增(RPA)是一种高度灵敏且具有选择性的等温扩增技术,在37-42°C下运行,样品制备要求极低,能够在不到20分钟的时间内扩增低至1-10个DNA目标拷贝。它已被用于扩增各种目标,包括来自多种生物体和样品的RNA、miRNA、单链DNA和双链DNA。越来越多详细介绍RPA应用的出版物不断涌现,扩增已在溶液相、固相中以及桥式扩增形式中进行。此外,RPA已成功与不同的检测策略相结合,从终点侧流试纸条到实时荧光检测等等。本综述重点关注与RPA技术相关的不同方法和进展,并突出该技术的一些优点和缺点。