Next-generation CRISPR-based molecular diagnostics for veterinary parasitology: From laboratory innovation to one health surveillance

Abstract

Veterinary parasitic diseases remain a major threat to livestock production, companion animal health, wildlife conservation, and public health worldwide. Conventional diagnostic methods, including microscopy, serology, and culture-based techniques, often suffer from limited sensitivity, poor specificity, prolonged turnaround time, and dependence on specialized laboratory facilities. Recent advances in Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based diagnostics have introduced highly sensitive, rapid, and field-deployable platforms for parasite detection. This review critically summarizes the principles and recent applications of CRISPR/Cas12, Cas13, and emerging Cas14-based diagnostic approaches in veterinary parasitology, covering protozoan, helminthic, and arthropod-borne parasites. It further compares CRISPR platforms and discusses their integration with isothermal amplification, lateral flow assays, fluorescence detection, and portable biosensors to improve diagnostic performance. Moreover, current challenges, including sample preparation, assay standardization, multiplex detection, regulatory approval, and commercialization, are highlighted. Finally, future perspectives on amplification-free CRISPR assays, artificial intelligence-assisted diagnostics, and point-of-care platforms are discussed. By providing a translational perspective from laboratory innovation to field application, this review highlights the potential of CRISPR-based diagnostics to transform veterinary parasite surveillance and disease control.