Abstract Rapid and sensitive detection of foodborne pathogens in food industry is of high importance in day-to-day practice to ensure safe food. To address this issue, multiple foodborne pathogens are targeted for rapid identification based in DNA amplification. A 3D PDMS sponge was fabricated using salt crystals as scarifying mold and functionalized with a ligand, apolipoprotein-H (ApoH), to test bacterial capturing for both Gram positive (L. monocytogenes) and negative bacteria (Salmonella spp.), in a microfluidic device. Pure culture of both pathogens in a range of ~10–105 CFU/mL were tested and the application of the developed automated pre-concentration protocol in real samples was verified using spiked surface samples after swab sampling. Bacterial DNA was extracted directly from the sponge and used for Real Time quantitative Polymerase Chain Reaction (qPCR) detection. The sponges did not show any significant resistance to sample flow and could easily be incorporated in a microfluidic device. A capture efficiency above 70% was observed for both targeted (Gram positive and Gram negative) pathogens and a Limit of Detection (LoD) in the range of 103 and 104 CFU/mL was obtained for Salmonella spp. and L. monocytogenes, respectively. Using this approached, we are able to perform multiplexed (Gram positive and Gram negative) capturing and reduce the enrichment time compared to the gold standard plate culture (over 1-day) method. The use of a 3D sponge for direct capturing of multiplexed pathogen on microfluidic device, followed by qPCR detection is an efficient and versatile method to stratify the presence of bacteria. This approach and methodology has potential to be integrated in full automatized device and used as point of need (PoN) system for foodborne pathogen stratification in food packaging/production industries.

中文翻译：

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用于快速多重检测食源性病原体的智能微流体平台

摘要 食品工业中食源性病原体的快速、灵敏检测对于确保食品安全在日常实践中具有重要意义。为了解决这个问题，多种食源性病原体成为基于 DNA 扩增的快速识别目标。3D PDMS 海绵是使用盐晶体作为划痕模具制造的，并用配体载脂蛋白-H (ApoH) 进行功能化，以在微流体中测试革兰氏阳性菌（单核细胞增生李斯特菌）和阴性菌（沙门氏菌属）的细菌捕获设备。测试了约 10–105 CFU/mL 范围内的两种病原体的纯培养物，并在拭子取样后使用加标表面样品验证了开发的自动预浓缩方案在实际样品中的应用。细菌 DNA 直接从海绵中提取，用于实时定量聚合酶链反应 (qPCR) 检测。海绵对样品流动没有任何明显的阻力，可以很容易地结合到微流体装置中。观察到目标（革兰氏阳性和革兰氏阴性）病原体的捕获效率均高于 70%，沙门氏菌的检测限 (LoD) 为 103 至 104 CFU/mL。和单核细胞增生李斯特菌，分别。使用这种方法，我们能够执行多重（革兰氏阳性和革兰氏阴性）捕获并减少与金标准板培养（超过 1 天）方法相比的富集时间。使用 3D 海绵在微流体装置上直接捕获多重病原体，其次是 qPCR 检测是一种有效且通用的方法来对细菌的存在进行分层。这种方法和方法有可能被集成到全自动设备中，并用作食品包装/生产行业中食源性病原体分层的需求点 (PoN) 系统。