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CRISPR-Cas12a-Mediated Hue-Recognition Lateral Flow Assay for Point-of-Need Detection of Salmonella

文献类型: 外文期刊

作者: Yuan, Jingrui 1 ; Wang, Liu 2 ; Huang, Liang 1 ; He, Kaiyu 2 ; Wang, Hongmei 2 ; Xu, Xiahong 2 ; Su, Bin 3 ; Wang, Jing 1 ;

作者机构: 1.Zhejiang Univ Technol, Coll Chem Engn, Hangzhou 310014, Peoples R China

2.Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, State Key Lab Managing Biot & Chem Threats Qual &, Key Lab Informat Traceabil Agr Prod,Minist Agr & R, Hangzhou 310021, Peoples R China

3.Zhejiang Univ, Inst Analyt Chem, Dept Chem, Hangzhou 310058, Peoples R China

期刊名称:ANALYTICAL CHEMISTRY ( 影响因子:7.4; 五年影响因子:7.0 )

ISSN: 0003-2700

年卷期: 2023 年 96 卷 1 期

页码:

收录情况: SCI

摘要: Nucleic acid detection of pathogens in a point-of-need (PON) manner is of great significance yet remains challenging for sensitive and accurate visual discrimination. Here, we report a CRISPR-Cas12a-mediated lateral flow assay for PON detection of Salmonella typhimurium (S.ty) that is a prevailing pathogen disseminated through tainted food. The variation of the fluorescence color of the test line is exploited to interpret the results, enabling the discrimination between positive and negative samples on the basis of a hue-recognition mechanism. By leveraging the cleavage activity of Cas12a and hue-recognition readout, the assay facilitated by recombinase polymerase amplification can yield a visual detection limit of 1 copy mu L-1 for S.ty genomic DNA within 1 h. The assay also displays a high specificity toward S.ty in fresh chicken samples, as well as a sensitivity 10-fold better than that of the commercial test strip. Moreover, a semiquantitative detection of S.ty ranging from 0 to 4 x 10(3) CFU/mL by the naked eye is made possible, thanks to the easily discernible color change of the test line. This approach provides an easy, rapid, accurate, and user-friendly solution for the PON detection of Salmonella and other pathogens.

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