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

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

Authors :
Li, Lina
Duan, Canxing
Weng, Jianfeng
Qi, Xiantao
Liu, Changlin
Li, Xinhai
Zhu, Jinjie
Xie, Chuanxiao
Source :
SCIENCE CHINA Life Sciences; July 2022, Vol. 65 Issue: 7 p1456-1465, 10p
Publication Year :
2022

Abstract

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 graminearumand Fusarium verticillioidesand 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.

Details

Language :
English
ISSN :
16747305 and 18691889
Volume :
65
Issue :
7
Database :
Supplemental Index
Journal :
SCIENCE CHINA Life Sciences
Publication Type :
Periodical
Accession number :
ejs58589927
Full Text :
https://doi.org/10.1007/s11427-021-2028-x