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Review
. 2025 Jul 30:15:1640938.
doi: 10.3389/fcimb.2025.1640938. eCollection 2025.

Research progress on the application of RPA-CRISPR/Cas12a in the rapid visual detection of pathogenic microorganisms

Tuo Ji  1   2   3 , Xin Fang  4 , Yuzhi Gao  1   2   3 , Kun Yu  1   2   3 , Xuzhu Gao  1   2   3
Affiliations
Review

Research progress on the application of RPA-CRISPR/Cas12a in the rapid visual detection of pathogenic microorganisms

Tuo Ji et al. Front Cell Infect Microbiol. .

Abstract

In an increasingly complex global public health landscape, the continuous emergence of novel pathogens and the growing problem of antibiotic resistance highlight the urgent need for rapid, efficient, and precise detection technologies for pathogenic microorganisms. The innovative combination of Recombinase Polymerase Amplification (RPA) and CRISPR/Cas12a enables the rapid amplification of target gene fragments under isothermal conditions and the precise recognition and cleavage of specific nucleic acid sequences. The integration of RPA and CRISPR/Cas12a significantly enhances the sensitivity and accuracy of detection simplifies operational procedures, and reduces the dependence on specialized equipment for testing personnel. This combination demonstrates great potential for application in clinical diagnostics and point-of-care testing. This article provides a detailed overview of the principles of RPA-CRISPR/Cas12a and its latest research progress in the field of pathogen detection, aiming to promote the widespread application of RPA-CRISPR/Cas12a technology in clinical medicine and public health and to offer theoretical support for its further optimization.

Keywords: CRISPR/Cas12a; RPA; pathogenic microorganisms; point-of-care testing; visualization.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Principle of RPA-CRISPR/Cas12a. Created in BioRender. Ji, (2025) https://BioRender.com/au4gcl0.
See this image and copyright information in PMC

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