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使用CRISPR-Cas13a与RT-RAA技术结合使用牛类冠状病毒的特定和敏感检测

Zili Liang¹, Ruxing Luo¹, Qifu He²

¹Key Laboratory of Animal Medicine at Southwest Minzu University of Sichuan Province, College of Animal Science and Veterinary Medicine, Southwest Minzu University, Chengdu, China.

Frontiers in veterinary science

|January 22, 2025

概括

使用CRISPR/Cas13a和RT-RAA技术的新分析方法可以准确检测牛冠状病毒 (BCoV). 这种方法为牛的早期疾病检测提供了更好的灵敏度和特异性,有助于疾病控制.

科学领域:

兽医病毒学兽医病毒学
分子诊断学分子诊断学
这就是CRISPR技术.

背景情况:

牛冠状病毒 (BCoV) 在牛中引起显著的肠道和呼吸道疾病,导致经济损失很大.
准确而敏感地检测BCoV对于有效的疾病预防和控制策略至关重要.

研究的目的:

开发和验证一种针对牛冠状病毒 (BCoV) 的特定,敏感和稳定的核酸检测试验.
为早期疾病预防和牛群中BCoV的流行病学监测提供一种新的工具.

主要方法:

将CRISPR/Cas13a技术与反转录复合酶辅助放大 (RT-RAA) 技术相结合.
针对RT-RAA和CRISPRRNA (crRNA) 的原体被设计为准BCoV.CoV的保存核体 (N) 基因.

主要成果:

该试验表明,BCoV.的低检测极限为1.72副本/μl.
没有观察到与其他10种常见的牛病原体的交叉反应.
该试验在临床样本中显示出高特异性和灵敏性,检测BCoV的阳性率为58.3%,明显高于RT-qPCR.
批量内和批量间的变化系数分别低于4.98%和4.58%,表明试验稳定性.

结论:

一个强大的RT-RAA-CRISPR/Cas13a测试用于BCoV检测已经成功开发.
这种测试为牛和雅克群体中BCoV的临床诊断和流行病学监测提供了增强的技术支持.

关键词:

这就是CRISPR/Cas13a.土耳其人土耳其人RT-RAAA 牛的冠状病毒病毒.诊断诊断诊断的诊断诊断诊断的诊断诊断的诊断核酸检测核酸检测检测

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