Recent advances in recombinase polymerase amplification: Principle, advantages, disadvantages and applications

Abstract

After the outbreak of SARS-CoV-2, nucleic acid testing quickly entered people’s lives. In addition to the polymerase chain reaction (PCR) which was commonly used in nucleic acid testing, isothermal amplification methods were also important nucleic acid testing methods. Among several common isothermal amplification methods like displaced amplification, rolling circle amplification, and so on, recombinase polymerase amplification (RPA) was recently paid more attention to. It had the advantages like a simple operation, fast amplification speed, and reaction at 37-42°C, et al. So it was very suitable for field detection. However, there were still some disadvantages to RPA. Herein, our review mainly summarized the principle, advantages, and disadvantages of RPA. The specific applications of RPA in bacterial detection, fungi detection, virus detection, parasite detection, drug resistance gene detection, genetically modified food detection, and SARS-CoV-2 detection were also described. It was hoped that the latest research progress on RPA could be better delivered to the readers who were interested in RPA.

Recombinase Polymerase Amplification Assay for the Field Detection of Mal Secco Disease byPlenodomus tracheiphilus

“…These methods differ from each other in terms of amplification temperatures, sensitivities, reaction times, and other advantages and drawbacks (Gill and Ghaemi, 2008; Ivanov et al ., 2021). More recently, diagnostic assays based on RPA have become widely used for the molecular-based diagnosis of diseases (Tan et al ., 2022). The outbreak of SARS-CoV-2 has further promoted the application of RPA in nucleic acid detection (Bai et al ., 2022).…”

“…The outbreak of SARS-CoV-2 has further promoted the application of RPA in nucleic acid detection (Bai et al ., 2022). Previous studies on RPA detection have shown several advantages of this technique, including its user-friendliness (in terms of simplicity of operations and low equipment requirements), high sensitivity, and great specificity (Tan et al ., 2022). Additionally, RPA best matches the requirement of ‘rapidity’.…”

Applicability of Smart Tools in Vegetable Disease Diagnostics

“…This technique has attracted great attention because it is insensitive to Taq polymerase inhibitors, and the DNA does not need to be isolated. Previous publications have reported the acceptable limit of detection of the assay [66,67]. Stringent optimization of reaction conditions is highly recommended, as inhibition may occur due to enzyme competition.…”

Application of CRISPR/Cas Technology in Biological Nano‐Analysis

“…The RPA technique requires amplification templates, primers and various components in addition to the two enzymes. [32] The process is probably that the recombinant enzyme protein forms a complex with the primers and then searches for the homologous sequence, and when it is found, it forms a d-loop structure to start the reaction. Micropropulsion reactor published by Tian et al based on Cas12a trans-cleavage activity and RPA studies.…”