CRISPR/Cas系统在寄生虫病诊断领域的研究进展

doi:10.16431/j.cnki.1671-7236.2025.06.033

摘要/Abstract

摘要： 多年来动物寄生虫病主要依靠传统镜检与PCR技术结合的方式进行诊断,在临床中确实起到了明显的防治效果，也控制了一些寄生虫病的流行。但寄生虫病多是慢性消耗性疾病，早期诊断尤为重要，而传统方法的灵敏度、简便性不能满足彻底控制寄生虫病的需求。因此，亟需灵敏度更高、便捷性更强的检测方法。近年来，CRISPR/Cas系统不断发展，已发现了多种Cas蛋白的功能，并广泛应用于寄生虫病原的核酸检测。研究人员针对第2类CRISPR/Cas系统中的Cas13与Cas12蛋白与重组酶聚合酶扩增(recombinase polymerase amplification，RPA)或PCR联用，筛选特异性靶标序列并设计crRNA，之后与Cas12、Cas13蛋白和荧光报告基团组成CRISPR/Cas核酸检测系统，可用于多种寄生虫病的诊断，具有快速、精准、便捷和廉价等特点，适用于现场检测，具有巨大的市场应用潜力。笔者就CRISPR/Cas系统的作用机制和分类，以及近年来其在寄生虫病核酸检测领域的研究进展进行综述，以期为寄生虫病的早期诊断提供参考依据。

关键词: CRISPR/Cas系统; Cas13; Cas12; crRNA; 核酸检测

Abstract: The detection of animal parasitic diseases has mainly relied on the combination of traditional microscopy and PCR technology for many years.It has indeed played a significant role in clinical practice,and has also controlled the prevalence of some parasitic diseases.However,parasitic diseases are mostly chronic consumptive diseases,and the significance of early diagnosis is particularly important.The sensitivity and simplicity of traditional methods cannot meet the needs of complete control of parasitic diseases.Therefore,the promotion of the use of more sensitive and more convenient detection methods are urgently needed.In recent years,the CRISPR/Cas system has been continuously developed,and a variety of Cas protein functions have been discovered and widely used in nucleic acid detection techniques for parasitic pathogens.The researchers screened the specific target sequence and designed crRNA for the combination of Cas13 protein and Cas12 proteins with recombinase polymerase amplification (RPA) or PCR in the type 2 CRISPR/Cas system,and then combined with Cas12,Cas13 proteins and fluorescent reporter group to form a CRISPR/Cas nucleic acid detection system,which can be used for the detection of various parasitic diseases.It has the characteristics of fast,accurate,convenient and cheap,and is suitable for on-site detection,and has great market application potential.In this paper,the mechanism and classification of CRISPR/Cas system and its research progress in the field of nucleic acid diagnosis of parasitic diseases in recent years are reviewed,so as to provide reference for the early diagnosis of parasitic diseases.

Key words: CRISPR/Cas system; Cas13; Cas12; crRNA; nucleic acid detection

中图分类号:

S961.6

刘浩, 张昊, 张艳敏, 张丽丽, 王文龙. CRISPR/Cas系统在寄生虫病诊断领域的研究进展[J]. 中国畜牧兽医, 2025, 52(6): 2809-2817.

LIU Hao, ZHANG Hao, ZHANG Yanmin, ZHANG Lili, WANG Wenlong. Research Progress on CRISPR/CAS System in the Diagnosis of Parasitic Diseases[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2809-2817.

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