A novel system with robust compatibility and stability for detecting Sugarcane yellow leaf virus based on CRISPR-Cas12a
文献类型： 外文期刊
第一作者： Wang, Ting
作者： Wang, Ting;Li, Anzhen;Zhao, Hong;Guo, Jinlong;Tian, Helei;Wang, Jingwen;Que, Youxiong;Xu, Liping;Wu, Qibin;Que, Youxiong
作者机构：
关键词： Sugarcane yellow leaf virus; MIRA; CRISPR-Cas12a; sugarcane
期刊名称：MICROBIOLOGY SPECTRUM （ 影响因子：3.7； 五年影响因子：4.2 ）
ISSN： 2165-0497
年卷期： 2024 年
页码：
收录情况： SCI
摘要： Sugarcane yellow leaf virus (SCYLV) can reduce sugarcane productivity. A novel detection system based on reverse transcription-multienzyme isothermal rapid amplification (RT-MIRA) combined with CRISPR-Cas12a, named RT-MIRA-CRISPR-Cas12a, was developed. This innovative approach employs crude leaf extract directly as the reaction template, streamlining the extraction process for simplicity and speed. Combining RT-MIRA and CRISPR-Cas12a in one reaction tube increases the ease of operation while reducing the risk of aerosol contamination. In addition, it exhibits sensitivity equivalent to qPCR, boasting a lower detection limit of 25 copies. Remarkably, the entire process, from sample extraction to reaction completion, requires only 52-57 minutes, just a thermostat water bath. The result can be observed and judged by the naked eye.
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