One-tube detection of Salmonella Typhimurium using LAMP and CRISPR-Cas12b

文献类型： 外文期刊

第一作者： Gong, Jiansen

作者： Gong, Jiansen;Jiang, Yi;Li, Tingting;Zhang, Di;Fu, Lixia;Dou, Xinhong

作者机构：

关键词： Salmonella Typhimurium; LAMP; CRISPR/Cas12b; rapid detection; one-tube; one-step

期刊名称：MICROBIOLOGY SPECTRUM （ 影响因子：3.7； 五年影响因子：4.2 ）

ISSN： 2165-0497

年卷期： 2024 年

页码：

收录情况： SCI

摘要： Salmonella enterica serovar Typhimurium (ST) is a predominant serovar causing foodborne illnesses worldwide. Traditional detection methods often face challenges, including the need for specialized equipment, skilled operators, and lengthy procedures. To address these limitations, we developed a rapid, sensitive, and specific ST detection method by integrating loop-mediated isothermal amplification (LAMP) with the clustered regularly interspaced short palindromic repeats and associated protein 12b (CRISPR/Cas12b) system, all within a single tube. Our results indicate that the LAMP-CRISPR/Cas12b reaction can be completed isothermally in under 1 h without requiring specialized instruments. The platform's limit of detection (LoD) is 12.5 copies per reaction. Additionally, the system demonstrated 100% inclusivity and exclusivity when tested against 30 reference strains, highlighting its specificity. In practical applications, the LoDs for ST in pure nucleic acid and contaminated fecal samples were 2.32 and 23.2 CFU/mL, respectively, with higher sensitivity observed in pure nucleic acid samples. Overall, our findings underscore the potential of the one-tube LAMP-CRISPR/Cas12b platform as a rapid, sensitive, and specific tool for ST detection, particularly in resource-limited settings.

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