Research progress of RPA-CRISPR/Cas12a in the detection of pathogenic microorganisms

Abstract:

Recombinase polymerase amplification (RPA) can efficiently amplify the target sequence at constant temperatures and is praised for high sensitivity, short time consumption, and low equipment dependence. Therefore, RPA is a widely used isothermal amplification technology in recent years. However, non-specific amplification of RPA has limited its further development. The clustered regularly interspaced short palindromic repeats/associated protein 12a (CRISPR/Cas12a) is capable of specifically cleaving the double strands and has similar optimal temperature to the PRA temperature. Therefore, researchers have combined RPA with CRISPR/Cas12a to improve the detection specificity. Moreover, this method demonstrates high sensitivity, a wide application range, and simple operation. Particularly, the RPA-CRISPR/ Cas12a-based approach shows a promising application prospect in the detection of pathogenic microorganisms. This paper introduces RPA-CRISPR/Cas12a regarding the principles, product detection strategies, and application, providing a reference for the further development and application of RPA-CRISPR/Cas12a in the detection of pathogenic microorganisms.