College of Food Science, Shenyang Agricultural University, Shenyang 110866, China.
Key Laboratory of Agricultural Information Acquisition Technology, Ministry of Agriculture and Rural Affairs, China Agricultural University, Beijing 100083, China.
Biosensors (Basel). 2022 Sep 5;12(9):726. doi: 10.3390/bios12090726.
Foodborne pathogenic bacteria have become a worldwide threat to human health, and rapid and sensitive bacterial detection methods are urgently needed. In this study, a facile microfluidic chip was developed and combined with recombinase-aided amplification (RAA) for rapid and sensitive detection of using a non-contact eddy heater for dynamic lysis of bacterial cells and a 3D-printed fan-shaped active mixer for continuous-flow mixing. First, the bacterial sample was injected into the chip to flow through the spiral channel coiling around an iron rod under an alternating electromagnetic field, resulting in the dynamic lysis of bacterial cells by this non-contact eddy heater to release their nucleic acids. After cooling to ~75 °C, these nucleic acids were continuous-flow mixed with magnetic silica beads using the fan-shaped mixer and captured in the separation chamber using a magnet. Finally, the captured nucleic acids were eluted by the eluent from the beads to flow into the detection chamber, followed by RAA detection of nucleic acids to determine the bacterial amount. Under the optimal conditions, this microfluidic chip was able to quantitatively detect from 1.1 × 10 to 1.1 × 10 CFU/mL in 40 min with a detection limit of 89 CFU/mL and might be prospective to offer a simple, low-cost, fast and specific bacterial detection technique for ensuring food safety.
食源性病原体已成为全球范围内人类健康的威胁,因此急需快速、灵敏的细菌检测方法。本研究开发了一种简便的微流控芯片,并结合重组酶辅助扩增(RAA),通过非接触式电涡流加热器对细菌细胞进行动态裂解,以及 3D 打印的扇形主动混合器进行连续流动混合,实现了对 的快速、灵敏检测。首先,将细菌样品注入芯片,在交变电磁场的作用下,流经围绕铁棒盘旋的螺旋通道,通过非接触式电涡流加热器实现细菌细胞的动态裂解,释放其核酸。冷却至~75°C 后,使用扇形混合器将这些核酸与磁性硅胶珠连续流动混合,并在分离室中用磁铁捕获。最后,通过洗脱液从珠上洗脱捕获的核酸,使其流入检测室,然后进行 RAA 检测核酸以确定细菌数量。在最佳条件下,该微流控芯片能够在 40 分钟内定量检测 1.1×10 至 1.1×10 CFU/mL 的浓度,检测限为 89 CFU/mL,有望为确保食品安全提供一种简单、低成本、快速和特异的细菌检测技术。