Laboratory of Applied Micro and Nanotechnology (LAMINATE), DTU-Bioengineering (Department of Biotechnology and Biomedicine), Technical University of Denmark, Kgs. Lyngby 2800, Denmark.
Biolabchip Group, DTU-Bioengineering (Department of Biotechnology and Biomedicine), Technical University of Denmark, Kgs. Lyngby 2800, Denmark.
Anal Chem. 2023 Aug 29;95(34):12656-12663. doi: 10.1021/acs.analchem.3c00970. Epub 2023 Aug 16.
Accurate and rapid detection of pathogens in foods of animal origin has been a critical part of the One Health Action Plan of the European Union (EU). Biosensors have the potential in bringing required technologies to accomplish this on the field, wherein loop-mediated isothermal amplification (LAMP) and lab-on-a-chip have proven to be ideal. We have developed a LAMP-based point-of-care (POC) device, the VETPOD, as a solution to the contemporary challenges in the rapid detection of spp. The core technology in the VETPOD is a ready-to-use cartridge that included an injection-molded polymer chip with pyramid-shaped optical structures embedded within the chip. These pyramid-shaped optical structures direct the incident light, due to total internal reflection (TIR), through the reaction chambers to the phototransistor. The VETPOD was validated against the ISO 6579-1 reference method. A total of 310 samples were tested that included 180 spiked samples in 6 different meat categories and 130 strains to determine the specificity. The overall results were satisfactory, wherein the VETPOD had an acceptable sensitivity (96.51%) compared to the reference (98.81%) and near perfect agreement with ISO 6579-1 with an overall Cohen's kappa of 0.94. The relative level of detection (RLOD) for the VETPOD was 1.38 CFU/25 g that was found to be 1.17 times higher than the reference. The VETPOD showed 98% precision for inclusivity and 100% precision for the exclusivity samples. The VETPOD proved as a useful alternative to detect spp. that can be adaptable to a broader spectrum of pathogens in future.
准确、快速地检测动物源食品中的病原体一直是欧盟（EU）“同一健康行动计划”的关键部分。生物传感器有可能为在现场完成这一任务提供所需的技术，其中环介导等温扩增（LAMP）和芯片实验室已被证明是理想的选择。我们开发了一种基于 LAMP 的即时检测（POC）设备，即 VETPOD，以解决快速检测 spp. 面临的当代挑战。VETPOD 的核心技术是一个即用型试剂盒，其中包括一个注塑聚合物芯片，芯片内嵌入了金字塔形光学结构。这些金字塔形光学结构由于全内反射（TIR），将入射光引导通过反应室到达光电晶体管。VETPOD 已针对 ISO 6579-1 参考方法进行了验证。共测试了 310 个样本，其中包括 6 种不同肉类中的 180 个添加样本和 130 个菌株，以确定特异性。总体结果令人满意，VETPOD 的灵敏度（96.51%）与参考（98.81%）相当，与 ISO 6579-1 的总体一致性良好，Cohen's kappa 值为 0.94。VETPOD 的相对检测水平（RLOD）为 1.38 CFU/25 g，比参考值高 1.17 倍。VETPOD 对包容性样本的精度为 98%，对排他性样本的精度为 100%。VETPOD 被证明是一种有用的替代方法，可以检测 spp. 并在未来适应更广泛的病原体谱。
