A rapid detection method for Escherichia coli O157:H7 and Salmonella Typhimurium in food based on the combination of loop-mediated isothermal amplification and hybridization chain reaction

文献类型： 外文期刊

第一作者： Zhu, Kunpeng

作者： Zhu, Kunpeng;Kong, Jiaqi;Lao, Fei;Zhao, Liang;Li, Hui;Zhu, Kunpeng;Kong, Jiaqi;Chen, Ailiang;Li, Hui

作者机构：

关键词： LAMP; HCR; Specific binding probes; E. coliS; Typhimurium

期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.0； 五年影响因子：5.7 ）

ISSN： 0003-2670

年卷期： 2025 年 1374 卷

页码：

收录情况： SCI

摘要： Foodborne pathogens present a significant threat to public health, highlighting the critical need for rapid, accurate, and highly sensitive detection methods. This study introduces a novel combined method, integrating loop-mediated isothermal amplification (LAMP) and hybridization chain reaction (HCR), for the simultaneous detection of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium. LAMP amplification was initially performed to amplify the gene Z3276 and invA, followed by a binding reaction with the specific sequence designed for each amplification product. The above binding products were subsequently captured by streptavidin-modified magnetic beads (SA-MB), initiating the HCR reaction on their surface, with the primers H2 labeled with FAM or VIC fluorophores. Finally, fluorescence signals were measured using a microplate reader for a rapid screening of target pathogens. This method showed a high sensitivity of detection and achieved the detection limit of 1-2 log CFU/mL for E. coli O157:H7, and 2-3 log CFU/mL for S. Typhimurium either in model system or in real food such as apple juice. Moreover, this method effectively reduced the whole detection time to less than 3 h through the optimization of the LAMP amplification, SA-MB enrichment and HCR signal amplification. The development of the combination usage of LAMP and HCR in this study can realize the direct detection of E. coli O157:H7 and S. Typhimurium in real food without the requirement of a pre-enrichment step, while effectively shortening the reaction time and thus satisfy the demand food safety supervision for rapid detection of foodborne pathogens.

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