RPA技术及其在食品检测中的应用

The Technology of Recombinase Polymerase Amplification(RPA) and Its Usage on Food Detection

重组酶聚合酶扩增技术(recombinase polymerase amplification,RPA)是以T4噬菌体的核酸复制机制为基本原理,模仿细胞内的核酸复制机制的一种新兴恒温核酸扩增技术。在多种酶的辅助下,31℃~37℃下恒温20 min即可完成核酸的体外扩增。它具有反应快速、高敏感性、高特异性、低设备依赖性等传统体外核酸扩增技术所不具备的优点。利用RPA技术可以在简易实验设备甚至野外条件下实现对核酸的快速扩增,结合侧流层析试纸技术或荧光检测装置即可对扩增结果进行快速定性或定量检测。但是,RPA技术作为一门新兴技术,在研究方面的应用鲜有提及,尤其在国内。本综述了RPA技术的原理、使用条件、优缺点及其在食品安全检测方面的应用,旨在为该技术今后的深入研究和应用提供一定的参考。

The technology of recombinase polymerase amplification （RPA） is an emerging isothermal nucleic acid amplification technology which is based on the mechanism of nucleic acid replication of T4 phages and mimicks the mechanism of nucleic acid replication in ceils. With the help of many enzymes, nucleic acid can be amplified in vitro at 31 ℃-37℃ in 20 minutes. It has many advantages which the traditional in vitro nucleic acid amplification technologies do not possess, such as rapid reaction, high sensitivity, high specificity and low dependence of equipment. RPA technology can be used in simple laboratory equipment and field conditions for the rapid amplification of nucleic acid. Combined with the technique of lateral flow chromatography paper or fluorescence detection device, a rapid qualitative or quantitative detection of the amplification results can be performed. However, as an emerging technology, the application of RPA technology on research is scarcely mentioned, especially in the domestic. In this review, we summarized the theory, working conditions, merits and faults of RPA technology and it＇s application in the food detection, aiming to provide some guidance for further development and application of this method.