重组酶聚合酶扩增:扩增原理及性能分析

Recombinase Polymerase Amplification:from Principle to Performance

脱氧核糖核酸(DNA)放大技术对于核酸检测(NAT)至关重要.聚合酶链式反应(PCR)虽然是核酸检测的基准扩增技术,但其主要适用于条件较好的中心实验室.重组酶聚合酶扩增(RPA)是一种非常有潜力的等温扩增技术,对仪器设备依赖性小,适用于资源贫乏地区.因此,该技术在核酸检测时不受实验场所限制,非常适合即时检测(POCT).作为一种正在快速发展的扩增技术,RPA也存在阻碍其进一步发展的缺陷.本文对RPA的扩增原理和扩增性能进行了总结,重点讨论了对扩增性能至关重要的引物重组和ATP动态平衡调控过程,并概述了RPA存在的缺陷和潜在的解决方向.

DNA amplification is essential to most nucleic acid testing(NAT).There is no doubt that poly⁃merase chain reaction(PCR)is the benchmark technology for DNA amplification,but is mainly performed in centralized laboratories.Recombinase polymerase amplification(RPA),as one of the most promising isother⁃mal amplification technologies,can break through laboratory boundaries for nucleic acid testing,since it is less instrument-dependent and easier access to resource-limited areas,namely,suitable for point-of-care testing(POCT)applications.As a developing technology,RPA also has several defects,which hind its wide⁃spread applications and development.Here,we make a summary on the principle and performance of RPA,with particular interest on the key process of the primer recombination and the dynamic balance of ATP that guide the eventual performance.Besides,several critical issues and potential points for further improving the performance of RPA were also summarized.