A portable microfluidic analyzer for integrated bacterial detection using visible loop-mediated amplification
A portable microfluidic analyzer for integrated bacterial detection using visible loop-mediated amplification
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DOI：
10.1016/j.snb.2020.127834
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发表时间：
2020-05-01
影响因子：
8.4
通讯作者：
Xu, Youchun
中科院分区：
文献类型：
--
作者：
Liu, Dongchen;Zhu, Yunzeng;Xu, Youchun
近年来，细菌污染已成为食品安全的重要问题。基于培养的方法虽然是细菌检测的金标准，但通常需要很长时间进行检测。基于微流控的分子诊断有助于实现细菌的快速、准确检测；然而，现有的微流体系统的便携性有限，因为需要笨重且复杂的仪器。在本研究中，开发了一种便携式分析仪，可在离心芯片上快速同时检测五种食源性细菌（金黄色葡萄球菌、沙门氏菌、志贺氏菌、产肠毒素大肠杆菌和铜绿假单胞菌）。包括细菌裂解、可见环介导扩增 (LAMP) 和检测在内的流程整合在 70 分钟内完成。该分析仪尺寸紧凑（151 x 134 x 110 mm），两个电机位于离心芯片两侧。上部电机用于产生旋转磁场，通过珠子敲打来裂解细菌，而下部电机则根据需要旋转芯片以执行以下过程：细菌裂解物的沉淀、LAMP缓冲液和细菌裂解物的混合以及将混合物分配到指定的反应室。 LAMP 结果由分析仪中的颜色传感器检测。使用这种便携式分析仪，与 Eppendorf 管中的传统 LAMP 相比，对五种细菌实现了相似的检测灵敏度。考虑到系统的便携性及其操作的简单性，本研究中开发的分析仪可能在护理点应用中具有潜在用途。
Bacterial contamination has become an important issue for food safety in recent years. Culture-based methods, though a gold standard for bacterial detection, usually take a long time for detection. Microfluidic-based molecular diagnosis can help to achieve rapid and accurate detection of bacteria; however, existing microfluidic systems have limited portability because bulky and complicated instruments are needed. In this study, a portable analyzer was developed to rapidly and simultaneously detect five types of foodborne bacteria (Staphylococcus aureus, Salmonella, Shigella, enterotoxigenic Escherichia coll., and Pseudomonas aeruginosa) on a centrifugal chip. Integration of the processes, including bacteria lysis, visible loop-mediated amplification (LAMP) and detection, was accomplished within 70 min. The analyzer has a compact size (151 x 134 x 110 mm) and two motors positioned on both sides of the centrifugal chip. The upper motor was used to generate a rotating magnetic field for lysing bacteria by bead-beating, whereas the lower motor rotated the chip on demand to execute the following processes: sedimentation of bacteria lysate, mixing of the LAMP buffer and bacteria lysate, and distribution of the mixture to the designated reaction chambers. The LAMP results were detected by a color sensor in the analyzer. Using this portable analyzer, similar detection sensitivities for the five bacteria were achieved compared to that of conventional LAMP in Eppendorf tubes. Considering the portability of the system and its simplicity of operation, the analyzer developed in this study may be of potential use in point-of-care applications.