生物
胡椒粉
病毒学
清脆的
重组酶聚合酶扩增
烟草病毒
反式激活crRNA
遗传学
植物病毒
病毒
计算生物学
生物技术
聚合酶链反应
园艺
基因
Cas9
作者
Zhenxing Zhao,Siyuan Wang,Zheng Dong,Qixuan Fan,Rong Lei,Ruirui Kuang,Yongjiang Zhang
标识
DOI:10.1021/acs.jafc.3c05268
摘要
Several tobamoviruses cause substantial economic losses to tomato and pepper crops globally, especially the pepper mild mosaic virus (PMMoV), tomato brown rugose fruit virus (ToBRFV), tomato mosaic virus (ToMV), and tomato mottle mosaic virus (ToMMV). A fast and accurate detection method is essential for virus identification. An all-in-one reaction method combining a one-step reverse-transcription recombinase-aided amplification (RT-RAA) and CRISPR/Cas12a-based lateral flow assay in one mixture was developed to rapidly screen and accurately differentiate among these four tobamoviruses for field detection in tomato and pepper plants. With a generic RT-RAA primer set and a mix of four specific crRNAs, along with a portable metal incubator and the use of a crude extraction method, this method screened for PMMoV, ToBRFV, ToMV, and ToMMV concurrently in less than 1 h, enabling field workers to take action immediately. The accurate differentiation of these four viruses could be achieved by later adding a single specific crRNA.
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