Current status of recombinase polymerase amplification technologies for the detection of pathogenic microorganisms

Rapid detection of pathogenic microorganisms is key to the epidemiologic identification, prevention and control of disease in the field of public health. PCR-based pathogen detection methods have been widely used because they overcome the time-consuming issues encountered in traditional culture-based methods, including the limited detecting window-phase of immunological detection. However, the requirement for precise temperaturecontrolled thermal cyclers severely limits the application of these methods in resource-limited areas. Recombinase polymerase amplification (RPA) is a new type of nucleic acid amplification technology that can amplify DNA or RNA at a constant temperature. It has the advantages of simple operation, high specificity and sensitivity and a short detection time. In recent years, a number of alternative methods for pathogenic microorganism detection have been developed by combining microfluidic technology with RPA. Through the design of chip structures, optimization of injection modes, and utilization of multiple detection and quantification methods, the integration of pathogen nucleic acid extraction, amplification and detection is achieved, and this approach is suitable for the rapid detection of pathogenic microorganisms in various environments. In this review, we compare different nucleic acid amplification techniques, explain the principle of RPA technology, detection methods, and applications for pathogen microorganism detection and describe future direction of RPA application. These methods increase the ability to rapidly screen pathogenic microorganisms, thus improving the management of infectious diseases in the field of public health.

Automated summary

Rapid detection of pathogenic microorganisms is crucial for epidemiologic identification, prevention, and control in public health. PCR-based methods have limitations in resource-limited areas due to the need for precise temperature control. Recombinase polymerase amplification (RPA) is a new nucleic acid amplification technology that can amplify DNA or RNA at a constant temperature, offering simplicity, high specificity and sensitivity, and short detection time. Microfluidic technology combined with RPA enables rapid detection of pathogenic microorganisms by integrating nucleic acid extraction, amplification, and detection.