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集成LAMP-CRISPR/Cas12a技术的便携式DNA提取用于鼠伤寒沙门氏菌的现场检测。

Portable DNA extraction integrated with LAMP-CRISPR/Cas12a technology for on-site detection of Salmonella Typhimurium.

作者信息

Zhang Liang, Zhang Mengfan, Liu Xingxing, Wei Jia, Yin Caihong, Wang Nan, Fan Beibei, Fu Yanli, Liu Yanwen, Bu Liangyun, Su Zhenyue, Pang Bo, Li Jinhua, Song Xiuling

机构信息

School of Public Health, Jilin University, 130021, Changchun, P.R. China.

Disease Control and Prevention of Liaoning Province, 110000, Shenyang, P.R. China.

出版信息

NPJ Sci Food. 2025 Mar 24;9(1):39. doi: 10.1038/s41538-025-00401-2.

Abstract

The infection and outbreak of Salmonella typhimurium (S. typhimurium) highlight the need for developing a reliable on-site detection strategy fitting to various settings. However, due to the requirement of specialized instruments and trained personnel, traditional detection methods have to be implemented in laboratories and are not ideal for on-site applications. To achieve a sample-to-answer and field-deployable detection for S. typhimurium, we developed an integrated nucleic acid detection platform combining single-vial of loop-mediated isothermal amplification (LAMP)-clustered regularly interspaced short palindromic repeat (CRISPR)/Cas12a system, portable device and smartphone app. This platform enables the extraction, concentration, and purification of DNA, amplification of the target, and output of visual fluorescent signals within 1 h. With this detection platform, 10 CFU/mL of S. typhimurium in food and environmental matrix was able to be accurately detected. This method demonstrated excellent selectivity. Also, an auxiliary smartphone application was developed to achieve simplified result interpretation. Our method exhibited potential to better control and respond to outbreaks of foodborne diseases, especially in low-resource settings.

摘要

鼠伤寒沙门氏菌(S. typhimurium)的感染和爆发凸显了开发适用于各种环境的可靠现场检测策略的必要性。然而,由于需要专业仪器和经过培训的人员,传统检测方法必须在实验室中实施,并不适合现场应用。为了实现对鼠伤寒沙门氏菌的样本到答案且可现场部署的检测,我们开发了一种集成核酸检测平台,该平台结合了单管环介导等温扩增(LAMP)-成簇规律间隔短回文重复序列(CRISPR)/Cas12a系统、便携式设备和智能手机应用程序。该平台能够在1小时内完成DNA的提取、浓缩和纯化、目标物的扩增以及视觉荧光信号的输出。利用这个检测平台,能够准确检测食品和环境基质中10 CFU/mL的鼠伤寒沙门氏菌。该方法显示出优异的选择性。此外,还开发了一个辅助智能手机应用程序以实现简化的结果解读。我们的方法在更好地控制和应对食源性疾病爆发方面展现出潜力,尤其是在资源匮乏的环境中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/74c3/11933393/4293617ca78e/41538_2025_401_Fig1_HTML.jpg

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