复合酶聚合酶放大与CRISPR/Cas12a技术相结合,用于快速现场检测腺病毒3
Qi-Zhang Liang1, Wei Chen1,2, Yuhai Bi3
1Fujian Academy of Agricultural Sciences/Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention, Institute of Animal Husbandry and Veterinary Medicine, Fuzhou, China.
Frontiers in microbiology
|August 22, 2025
概括
使用RPA和CRISPR/Cas12a技术开发了一种新的腺病毒3 (DAdV-3) 快速检测方法. 这种易于使用的测定方法可准确地在现场检测DAdV-3,对于控制致命的病至关重要.
科学领域:
- 兽医病毒学
- 分子诊断
- 鸟类病理学
背景情况:
- 腺病毒3 (DAdV-3) 在子中引起严重的肝损伤和出血,导致高发病率和死亡率.
- 同时感染和缺乏有效的DAdV-3疫苗/治疗对养业构成重大挑战.
- 准确和快速的诊断工具对于及时干预和疾病管理至关重要.
研究的目的:
- 开发一种快速,特定和敏感的腺病毒3 (DAdV-3) 检测方法.
- 在不需要复杂的实验室设备的情况下,能够在现场检测DAdV-3.
主要方法:
- 开发了与重组酶聚合酶放大 (RPA) 和CRISPR/Cas12a技术相结合的侧向流带 (LFS) 试验.
- 针对DAdV-3纤维-2基因进行特定的检测.
- 使用95个水鸟样本进行验证,并与定量聚合酶链反应 (qPCR) 进行比较.
主要成果:
- RPA-CRISPR/Cas12a-LFS方法实现了低检测极限为3.0个基因拷贝.
- 具有很高的特异性,与其他八种禽类病毒没有交叉反应.
- 在分析水禽样本时,达到98.95%的一致性和与qPCR的一致性,总反应时间为37°C的45分钟.
结论:
- 开发的RPA-CRISPR/Cas12a-LFS方法是一种用户友好,快速,灵敏和准确的现场DAdV-3检测工具.
- 这种测试具有在子行业有效预防和控制策略的巨大潜力.
相关概念视频
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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...


