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基于CRISPR-Cas13a系统,为禽流感病毒建立快速视觉检测方法

Zongshu Zhang¹, Chunguang Wang¹, Xi Chen¹

¹College of Veterinary Medicine, Hebei Agricultural University, Baoding, China.

Frontiers in veterinary science

|January 23, 2024

概括

使用复合酶辅助放大 (RAA) 和CRISPR-Cas13a技术的新视觉检测方法提供了快速,特定和敏感的禽流感病毒 (AIV) 检测. 这种RAA-CRISPR-Cas13a-LFD测定对AIV诊断具有高度敏感性和准确性.

科学领域:

分子生物学分子生物学
病毒学病毒学
生物技术是生物技术.

背景情况:

禽流感病毒 (AIV) 对家禽健康和全球粮食安全构成重大威胁.
快速,具体和敏感的诊断工具对于有效的AIV监测和控制至关重要.
现有的方法可能缺乏现场应用所需的速度,灵敏度或视觉解释性.

研究的目的:

开发一种针对禽流感病毒 (AIV) 的新型视觉检测方法.
建立一种结合复合酶辅助放大 (RAA) 与CRISPR-Cas13a系统的诊断试验.
评估新的RAA-CRISPR-Cas13a-LFD测定方法的特异性,敏感性和临床适用性.

主要方法:

设计的特定原料和CRISPRRNA (crRNA) 针对AIV.的保存核蛋白 (NP) 基因.
利用RAA技术快速放大目标基因片段.
使用CRISPR-Cas13a系统进行特定检测和侧流量计 (LFD) 进行视觉读取.

主要成果:

通过RAA-CRISPR-Cas13a-LFD方法证明了AIV的特定放大和视觉检测.
试验没有显示与其他常见的禽类病毒 (如IBV,ILTV和NDV) 的交叉反应.
获得了10^0副本/μL的高灵敏度,比PCR-糖电泳性能优于1000倍,临床样本的巧合率为98.75%,总反应时间约为1小时.

结论:

开发的RAA-CRISPR-Cas13a-LFD方法是一种快速,简单,特定和高度敏感的视觉诊断工具,用于AIV.
这种测定为及时和准确的AIV检测提供了有价值的新选择,有助于疾病管理.
测试的视觉性质增强了其在资源有限的环境或护理点诊断中使用的潜力.

关键词:

与CRISPR相关的蛋白质.聚类有规律间隔的短palindromic重复禽流感病毒是禽流感病毒的一种.侧向流量测量棒横向流量测量棒核酸检测核酸检测检测再组合酶辅助的放大.

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