Analytica chimica acta · 2025
Multi-target microfluidic loop-mediated isothermal amplification technology for the detection of foodborne pathogens in various food matrices.
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Lv Ji, Liu Wenping, Wang Fangfang, Xiong Lei, Yang Huabing, Yao Xiaowei, Liu Hongtao, Zheng Junping
Аннотация
BACKGROUND: Nutritional foods are susceptible to foodborne pathogens. Loop-mediated isothermal amplification (LAMP) technology has great potential in detecting foodborne pathogens, but conventional LAMP suffers from drawbacks such as complex operation and low detection throughput. Therefore, there is an urgent need to develop a multi-targeted and simultaneous LAMP detection platform to identify pathogens across various food matrices, in order to effectively manage food quality and safeguard human health.
RESULTS: The integration of LAMP with a microfluidic chip is proposed to enable the simultaneous and rapid detection of four pathogenic bacteria across eight different food matrices. We focused on designing and screening specific LAMP primers for Vibrio parahaemolyticus, Escherichia coli O157: H7, Yersinia enterocolitica, and Bacillus cereus. Then, we employed 10-well microfluidic chips to improve detection efficiency and reduce interference. We tested the feasibility and accuracy of detecting all four pathogens simultaneously on this chip platform. Our findings showed that the microfluidic LAMP detection platform could successfully identify the four pathogens with a low LOD (from 1.78 × 104 to 1.59 × 105 copies/mL) within 45 min. Furthermore, the platform specifically detected pathogens in a wide range of artificially simulated contaminated food matrices. All of these results could be observed with naked eyes and analyzed using fluorescence.
SIGNIFICANCE AND NOVELTY: The microfluidic LAMP platform, characterized by isothermal reactions, visualization features, robust immunity, and high throughput, presents an efficient, user-friendly, and highly sensitive approach for on-site detection. This technology has significant potential for future applications in the field of food safety monitoring.
RESULTS: The integration of LAMP with a microfluidic chip is proposed to enable the simultaneous and rapid detection of four pathogenic bacteria across eight different food matrices. We focused on designing and screening specific LAMP primers for Vibrio parahaemolyticus, Escherichia coli O157: H7, Yersinia enterocolitica, and Bacillus cereus. Then, we employed 10-well microfluidic chips to improve detection efficiency and reduce interference. We tested the feasibility and accuracy of detecting all four pathogens simultaneously on this chip platform. Our findings showed that the microfluidic LAMP detection platform could successfully identify the four pathogens with a low LOD (from 1.78 × 104 to 1.59 × 105 copies/mL) within 45 min. Furthermore, the platform specifically detected pathogens in a wide range of artificially simulated contaminated food matrices. All of these results could be observed with naked eyes and analyzed using fluorescence.
SIGNIFICANCE AND NOVELTY: The microfluidic LAMP platform, characterized by isothermal reactions, visualization features, robust immunity, and high throughput, presents an efficient, user-friendly, and highly sensitive approach for on-site detection. This technology has significant potential for future applications in the field of food safety monitoring.