动物疫病不仅严重危害动物健康，阻碍养殖业良性发展，而且对人类健康构成潜在威胁。开发灵敏、特异、快速的诊断方法对于疫病防控尤为重要。PCR是目前检测病原体核酸的金标准，但是它存在检测周期较长，需要精密仪器和专业技术人员，无法应用于现场即时检测等局限性。成簇规律间隔短回文重复序列及其相关蛋白(clustered regularly interspaced short palindromic repeats/CRISPR-associated protein，CRISPR/Cas)具有敏感性高、特异性强和操作简便等优点，可为动物疫病诊断提供新的方法和契机。CRISPR-Cas9靶向核酸的特异性和CRISPR-Cas12/Cas13"附属切割"活性使其在核酸检测中显示出巨大的应用前景。本文简述了 CRISPR/Cas系统的分类，对3种常用的CRISPR/Cas系统的核酸检测策略进行系统阐述，并对其在动物疫病诊断中应用的最新研究进展进行综述，最后讨论了它的优势、面临的挑战及未来发展方向，以期为动物疫病诊断新方法的开发提供参考。
Application of CRISPR/Cas-based Biosensors for Animal Diseases Diagnosis
Animal diseases not only seriously jeopardize the health of animals and hinder the healthy development of the breeding industry,but also pose a potential threat to human health.Therefore,it is vital to develop sensitive,specific and rapid diagnostic methods for disease prevention and control.The current gold standard for pathogens detection,PCR,has limitations such as long operational cycles,high instrumentation and technician requirements,cannot be applied to point-of-care testing.Clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeats-associated protein(CRISPR/Cas)have the advantages of high sensitivity,high specificity and easy operation,which can provide a new method and opportunity for the diagnosis of animal diseases.CRISPR-Cas9 specificity in targeting nucleic acids and"collateral cleavage"activity of CRISPR-Cas12/Cas13 show significant promise in nucleic acid detection.In this review,the classification of CRISPR/Cas systems,the strategies adopted by three commonly used CRISPR/Cas systems applied to nucleic acid detection were summarized,as well as the latest research progress of their application in animal disease diagnosis,finally their advantages,challenges,and future development were analyzed,in the hope of providing a reference for the application of CRISPR/Cas biosensing platforms in animal diseases diagnosis.
国家科技期刊平台
NSTL
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