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
作者： Wang, Ting 1 ; Li, Anzhen 1 ; Zhao, Hong 1 ; Wu, Qibin 2 ; Guo, Jinlong 1 ; Tian, Helei 1 ; Wang, Jingwen 1 ; Que, Youxiong 1 ; Xu, Liping 1 ;
作者机构： 1.Fujian Agr & Forestry Univ, Natl Engn Res Ctr Sugarcane, Key Lab Sugarcane Biol & Genet Breeding, Minist Agr & Rural Affairs,Coll Agr, Fuzhou, Peoples R China
2.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Natl Key Lab Trop Crop Breeding, Sanya, Peoples R China
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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