文献类型：期刊文献

英文题名：A lateral flow strip combined with Cas9 nickase-triggered amplification reaction for dual food-borne pathogen detection

作者：Wang, Luying[1];Shen, Xingying[1];Wang, Ting[2];Chen, Pinru[1];Qi, Nan[1];Yin, Bin-Cheng[1,2];Ye, Bang-Ce[1,2]

机构：[1]Zhejiang Univ Technol, Coll Pharmaceut Sci, Collaborat Innovat Ctr Yangtze River Delta Reg Gr, Inst Engn Biol & Hlth, Hangzhou 310014, Zhejiang, Peoples R China;[2]East China Univ Sci & Technol, Lab Biosyst & Microanal, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2020

卷号：165

外文期刊名：BIOSENSORS & BIOELECTRONICS

收录：;EI(收录号：20203008978027);WOS:【SCI-EXPANDED(收录号:WOS:000566442000018)】；

基金：This work was supported by grants from the National Key Research and Development Program of China (2018YFA0900404), National Natural Science Foundation of China (31730004, 21822402, 21675052) and China Postdoctoral Science Foundation (2019M652136).

语种：英文

外文关键词：Lateral flow strip; Cas9 nickase-based amplification reaction; Isothermal DNA amplification; Dual food-borne pathogen detection

摘要：Nucleic acid-based detection methods are accurate and rapid, which are widely-used in food-borne pathogen detection. However, traditional nucleic acid-based detection methods usually rely on special instruments, weakening their practicality for on-site tests in resource-limited locations. In this work, we developed a convenient and affordable method for food-borne pathogen detection based on a lateral flow strip combined with Cas9 nickase-triggered isothermal DNA amplification, which allows instrument-free and dual target detection. The genomic DNAs of two most common foodborne pathogens, Salmonella typhimurium and Escherichia coli, were simultaneously amplified in a one-pot reaction using specific sgRNAs and primers. The amplicons of genomic DNAs were double-labelled by digoxin/biotin and FITC/biotin tags, respectively, and directly visualized on a simple lateral flow strip. Our method exhibited a high specificity and sensitivity with a detection limit of 100 copies for genomic DNAs and 100 CFU/mL for bacteria. We believe that this method has potential to provide a convenient and low-cost point-of-care test for pathogen detection in the food quality surveillance.