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通过RPA-CRISPR/Cas12a调度的同热放大,用于快速检测Phytopythium化

Tingyan Xu¹, Fuliang Cao¹, Tingting Dai¹

¹Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu 210037, China.

|July 25, 2024

概括

一种新的重组酶聚合酶放大 (RPA) 和CRISPR/CRISPR相关蛋白12a (Cas12a) 测定方法可以快速检测隔离病原体Phytopythium helicoides. 这种敏感的方法有助于保护城市树木免受棕色腐烂等疾病的影响.

科学领域:

植物病理学植物病理学
分子诊断学分子诊断学
生物技术是生物技术.

背景情况:

植物 (Phytopythium helicoides) 是一种检疫病原体,对城市树木造成重大损害.
准确和快速的疾病诊断对于有效管理P.helicoides.是必不可少的.

研究的目的:

开发一种对Phytopythium helicoides的敏感和高效的检测方法.
使得早期诊断能够及时干预和控制疾病.

主要方法:

使用复合酶聚合酶放大 (RPA) 与CRISPR/CRISPR相关蛋白12a (Cas12a) 系统相结合.
优化检测条件,以实现高特异性和灵敏度.
验证了对人工注射植物组织的测定.

主要成果:

在RPA-CRISPR/Cas12a试验中,只检测到10 pg/μl的P. helicoides基因组DNA.
在37°C的20分钟内实现了检测,结果通过蓝光激发可视化.
该试验在识别 rhododendron 叶中的P. helicoides方面表现出高的特异性和灵敏性.

结论:

开发的RPA-CRISPR/Cas12a测定是一种敏感,高效和方便的工具,用于检测P.helicoides.
这种测试所促进的早期检测对于保护城市绿地覆盖物种至关重要.
这种方法为植物卫生管理中的检疫病原体诊断提供了有前途的方法.

关键词:

这就是CRISPR/Cas12a.植物的化物化物.检测检测检测检测检测疾病诊断疾病诊断复合酶聚合酶放大复合酶.

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