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使用CRISPR/Cas12a DETECTR试验结合异热复合酶聚合酶放大剂的快速和灵敏检测植物质体疾病

Wei Wei¹, Yinong Yang², Justin Shih³

¹Molecular Plant Pathology Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD, USA. wei.wei@usda.gov.

Methods in molecular biology (Clifton, N.J.)

|January 13, 2026

概括

这项研究介绍了一种新的DNA内核酶向CRISPR转载记者 (DETECTR) 试验,用于快速和具体检测植物细胞质植物疾病. 该试验结合了CRISPR/Cas12a技术与异热复合酶聚合酶放大 (RPA) 技术,用于敏感病原体的识别.

科学领域:

植物病理学植物病理学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

植物等离子体疾病对全球农业构成重大威胁.
目前的诊断方法可能是缓慢的,劳动密集的,或缺乏特异性.
需要快速,灵敏和现场部署的诊断工具.

研究的目的:

开发和介绍DNA内核酶向CRISPR转载记者 (DETECTR) 试验的详细方案.
为了快速和具体地检测植物中的植物质体疾病.
为研究人员和农业专业人员提供一个用户友好的方法.

主要方法:

使用了CRISPR/Cas12a技术与异热复合酶聚合酶放大 (RPA) 相结合.
RPA从植物基因组DNA中放大了目标植物等离子体DNA.
通过目标识别激活的Cas12a核酶将光报道器切割为信号生成.

主要成果:

DETECTR试验证明了植物质DNA的快速和特定检测.
该方法通过Cas12a介导的附带分离实现了高灵敏度.
提供了详细的样本准备,测试设置和数据解释指令.

结论:

DETECTR测定提供了一种先进和有效的诊断技术,用于检测植物质体.
该协议有助于采用CRISPR/Cas12a和RPA用于植物疾病诊断.
该试验有可能在农业研究和实践中得到广泛应用.

关键词:

克里斯普尔RNAs (克里斯普尔RNAs) (crRNAs) 是一种基因组.可以部署到现场.侧流免疫条纹侧流免疫条纹原生空间器相邻动图 (PAM)

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相关概念视频

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...