A field-deployable method for single and multiplex detection of DNA or RNA from pathogens using Cas12 and Cas13

文献类型： 外文期刊

第一作者： Li, Lina

作者： Li, Lina;Duan, Canxing;Weng, Jianfeng;Qi, Xiantao;Liu, Changlin;Li, Xinhai;Zhu, Jinjie;Xie, Chuanxiao;Li, Xinhai

作者机构：

关键词： nucleic acid detection; AsCas12a; LbCas12a; LwaCas13a; RfxCas13d; maize ear rot; Fusarium head blight; rice black-streaked dwarf virus (RBSDV)

期刊名称：SCIENCE CHINA-LIFE SCIENCES （ 影响因子：10.372； 五年影响因子：7.587 ）

ISSN： 1674-7305

年卷期： 2022 年 65 卷 7 期

页码：

收录情况： SCI

摘要： For some Cas nucleases, trans-cleavage activity triggered by CRISPR/Cas-mediated cis-cleavage upon target nucleic acid recognition has been explored for diagnostic detection. Portable single and multiplex nucleic acid-based detection is needed for crop pathogen management in agriculture. Here, we harnessed and characterized RfxCas13d as an additional CRISPR/Cas nucleic acid detection tool. We systematically characterized AsCas12a, LbCas12a, LwaCas13a, and RfxCas13d combined with isothermal amplification to develop a CRISPR/Cas nucleic acid-based tool for single or multiplex pathogen detection. Our data indicated that sufficient detection sensitivity was achieved with just a few copies of DNA/RNA targets as input. Using this tool, we successfully detected DNA from Fusarium graminearum and Fusarium verticillioides and RNA from rice black-streaked dwarf virus in crude extracts prepared in the field. Our method, from sample preparation to result readout, could be rapidly and easily deployed in the field. This system could be extended to other crop pathogens, including those that currently lack a detection method and have metabolite profiles that make detection challenging. This nucleic acid detection system could also be used for single-nucleotide polymorphism genotyping, transgene detection, and qualitative detection of gene expression in the field.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序