Improving food safety is crucial in the context of a “One Health” approach. To guarantee product quality and safety, the food industry, which has a very high turnover rate, needs short time-to-result analyses. Therefore, user-friendly systems at the point-of-need are necessary, presenting relevant analytical information and fulfilling the current regulations. To answer these challenges, a microfluidic platform integrating sample preparation and subsequent multiplex qPCR detection has been developed for on-site testing. The system consists of a fully automated instrument driving a microfluidic cartridge dedicated to the detection of multiple allergens in complex food matrices. The first part of the microfluidic cartridge contains pumps, reservoirs, valves and a filter to achieve DNA extraction, concentration and purification. Multiplex qPCR detection is carried out in the second part of the cartridge including a negative control chamber and five chambers for target analyte detection. The in-house developed instrument contains all functions to autonomously drive the microfluidic cartridge: pneumatic control for fluid actuation, thermal control for qPCR amplification and an optical system using three fluorescent wavelengths for multiplex detection of the target analytes and controls. We demonstrate the simultaneous detection of four different allergens – gluten, sesame, soy and hazelnut – from various complex food matrices. The turn-around-time from sample to result is close to two hours and controls in place validate the obtained results. For gluten, a direct comparison with ELISA shows that the regulatory threshold of 20 ppm is comfortably fulfilled. Moreover, all results are in agreement with external laboratory analyses performed in parallel on the same samples. Our findings confirm that the system can be used safely on-site without the risk of cross contamination between the various samples being analysed. In conclusion, our microfluidic platform offers a robust method for on-site allergen management.

中文翻译：

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用于现场 qPCR 分析的集成微流控平台：从样品到结果的食物过敏原检测


在“同一个健康”方法的背景下，改善食品安全至关重要。为了保证产品质量和安全，流动率非常高的食品行业需要较短的时间获得结果分析。因此，需要在需要时使用用户友好的系统，以提供相关的分析信息并满足当前法规。为了应对这些挑战，我们开发了一个集成样品制备和后续多重 qPCR 检测的微流控平台，用于现场检测。该系统由一台驱动微流体卡盒的全自动仪器组成，专门用于检测复杂食品基质中的多种过敏原。微流体卡盒的第一部分包含泵、储液槽、阀门和过滤器，以实现 DNA 提取、浓缩和纯化。多重 qPCR 检测在小柱的第二部分进行，包括一个阴性对照室和五个用于目标分析物检测的腔室。内部开发的仪器包含自主驱动微流体小柱的所有功能：用于流体驱动的气动控制、用于 qPCR 扩增的热控制以及使用三个荧光波长对目标分析物和对照进行多重检测的光学系统。我们演示了从各种复杂食品基质中同时检测四种不同的过敏原 - 麸质、芝麻、大豆和榛子。从样品到结果的周转时间接近 2 小时，并且适当的控制措施可以验证获得的结果。对于麸质，与 ELISA 的直接比较表明，可以轻松满足 20 ppm 的监管阈值。此外，所有结果都与对相同样品平行进行的外部实验室分析一致。 我们的研究结果证实，该系统可以在现场安全使用，而不会有被分析的各种样品之间交叉污染的风险。总之，我们的微流控平台为现场过敏原管理提供了一种强大的方法。