Molecular Physiology Laboratory, Cluster for Pioneering Research, RIKEN, Saitama 351-0198, Japan.
Anal Chem. 2023 Jun 27;95(25):9680-9686. doi: 10.1021/acs.analchem.3c01691. Epub 2023 Jun 12.
Genetic tests are highly sensitive, and quantitative methods for diagnosing human viral infections, including COVID-19, are also being used to diagnose plant diseases in various agricultural settings. Conventional genetic tests for plant viruses are mostly based on methods that require purification and amplification of viral genomes from plant samples, which generally take several hours in total, making it difficult to use them in rapid detection at point-of-care testing (POCT). In this study, we developed Direct-SATORI, a rapid and robust genetic test that eliminates the purification and amplification processes of viral genomes by extending the recently developed amplification-free digital RNA detection platform called SATORI, allowing the detection of various plant viral genes in a total of less than 15 min with a limit of detection (LoD) of 98 ∼ copies/μL using tomato viruses as an example. In addition, the platform can simultaneously detect eight plant viruses directly from ∼1 mg of tomato leaves with a sensitivity of 96% and a specificity of 99%. Direct-SATORI can be applied to various infections related to RNA viruses, and its practical use is highly anticipated as a versatile platform for plant disease diagnostics in the future.
遗传测试具有高度敏感性，用于诊断人类病毒感染的定量方法，包括 COVID-19，也被用于在各种农业环境中诊断植物疾病。传统的植物病毒遗传测试大多基于需要从植物样本中纯化和扩增病毒基因组的方法，这些方法通常总共需要几个小时，因此难以在即时护理检测（POCT）中进行快速检测。在这项研究中，我们开发了 Direct-SATORI，这是一种快速而强大的遗传测试，通过扩展最近开发的无扩增数字 RNA 检测平台 SATORI，消除了病毒基因组的纯化和扩增过程，允许在总共不到 15 分钟的时间内检测各种植物病毒基因，番茄病毒的检测限（LoD）为 98∼拷贝/μL。此外，该平台可以直接从约 1 毫克的番茄叶片中同时检测 8 种植物病毒，灵敏度为 96%，特异性为 99%。Direct-SATORI 可应用于与 RNA 病毒相关的各种感染，作为未来植物疾病诊断的通用平台，其实际应用具有很高的预期。
