Research progress of Class 2 CRISPR-Cas system in nucleic acid detection of animal pathogens

Abstract

The rapid spread of animal diseases and the evolution of associated pathogens underscore the urgent need for improved diagnostic techniques. Established nucleic acid detection methods typically rely on expensive and complex machinery, which requires specialized expertise and is time-consuming to operate. As a result, these methods are not well-suited for the monitoring and preliminary screening of epidemics in highly-intensive livestock operations. Therefore, there is a pressing need for the development of on-site rapid nucleic acid detection technologies that offer both high sensitivity and specificity. The clustered regularly interspaced short palindromic repeats and associated proteins (CRISPR-Cas) system is notable for its simplicity, precision and high-efficiency gene-editing capabilities. Recent investigations into CRISPR-Cas-based nucleic acid detection methods have demonstrated considerable potential for advancing diagnostic technology in this field. This paper provides a comprehensive review of CRISPR-Cas-based nucleic acid detection principles and their application in diagnosing animal diseases. It aims to serve as a valuable reference for researchers and practitioners involved in the development and implementation of CRISPR-Cas technologies for animal pathogen detection.

Graphical abstract

Keywords

CRISPR-Cas / nucleic acid detection / diagnosis / livestock

Highlight

	● Novel overview of Class 2 CRISPR-Cas systems for veterinary diagnostics.
	● Comparative analysis of Cas9, Cas12, and Cas13 detection platforms.
	● Integration with isothermal amplification enables rapid, ultrasensitive assays.
	● Multiplex readouts improve on-site applicability and diagnostic accuracy.
	● Future directions highlight amplification-free and field-deployable CRISPR tools.

RIGHTS & PERMISSIONS

The Author(s) 2025. Published by Higher Education Press. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0)