清脆的
化学
环介导等温扩增
STX2
大肠杆菌
检出限
反式激活crRNA
微生物学
色谱法
生物
基因
回文
志贺毒素
DNA
生物化学
作者
So‐Young Lee,Se‐Wook Oh
出处
期刊:Talanta
[Elsevier]
日期:2022-05-01
卷期号:241: 123186-123186
被引量:49
标识
DOI:10.1016/j.talanta.2021.123186
摘要
Escherichia coli O157:H7 is a major cause of fresh vegetable-associated infections that can threaten human health. A method for rapidly detecting food-borne pathogens should be developed for safe food management. A clustered regularly interspaced short palindromic repeats (CRISPR)-based detection method has the potential to greatly advance biosensing technology through its high sensitivity and specificity. In this study, we developed a rapid, sensitive, and visualized method of detecting E. coli O157:H7 (stx2 gene) based on a loop-mediated isothermal amplification (LAMP)-CRISPR/Cas12a system. The developed method was able to rectify the common false-negative results produced by LAMP, and the detection limit was 1.22 × 100 CFU/mL in pure culture. Furthermore, the LAMP-CRISPR/Cas12a system using filtration enrichment successfully detected 4.80 × 100 CFU/g of E. coli O157:H7 in romaine lettuce without pre-microbial enrichment culture. Consequently, the LAMP-CRISPR/Cas12a system is a useful technique for rapid and sensitive detection of E. coli O157:H7 in fresh products.
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