Identification for a Large-volume Food-borne Bacteria on a Fully Integrated Portable Centrifugal Disc

Hau Van Nguyen, V. D. Nguyen, E. Lee, T. Seo
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Abstract

Herein, we present a fully integrated portable centrifugal microsystem for multiplex detection of food poisoning bacteria with a large volume of sample up to 1 mL. The microsystem consists of a portable rotary genetic analyzer and a fully integrated lab-on-a-disc device. The portable rotary genetic analyzer is equipped with a couple of heating blocks, a motor and a UV-Vis optical detector. The device was designed with two units: a 3D printed solution-loading cartridge and a centrifugal microfluidic disc. All the essential solutions for the LAMP reaction (a sample, a washing, an elution and a LAMP cocktail solution) are stored inside the cartridge, and orderly released into centrifugal microdevice by a rotation program. Each unit of the device was designed with 20 reaction chambers for simultaneously detecting 19 kinds of food poisoning bacteria in one test. To increase the amount of a sample to 1 mL, we incorporated the super absorbent polymer (SAP) in the waste chamber to absorb the sample and washing solution during the device operation. The whole process was automatically conducted from bead-based DNA extraction to isothermal DNA amplification by LAMP reaction to colorimetric and UV-vis detection of amplicons in 60 min to identify three kinds of bacteria (Escherichia coli O157:H7, Salmonella Typhimurium, and Vibrio parahaemolyticus).
全集成便携式离心圆盘上大容量食源性细菌的鉴定
在此,我们提出了一种完全集成的便携式离心微系统,用于食物中毒细菌的多重检测,样品容量可达1ml。该微系统由便携式旋转遗传分析仪和完全集成的光盘实验室设备组成。便携式旋转基因分析仪配备了一对加热块,一个电机和一个紫外-可见光学探测器。该装置设计了两个单元:一个3D打印的溶液加载盒和一个离心微流控盘。LAMP反应的所有必要溶液(样品,洗涤,洗脱和LAMP鸡尾酒溶液)都储存在药筒内,并通过旋转程序有序地释放到离心微装置中。该装置每单元设计20个反应室,可同时检测19种食物中毒细菌。为了将样品的量增加到1ml,我们在废室中加入高吸水性聚合物(SAP),以在设备运行期间吸收样品和洗涤液。从珠状DNA提取到LAMP反应等温DNA扩增,再到比色法和紫外-可见法检测扩增产物,60 min内自动完成3种细菌(大肠杆菌O157:H7、鼠伤寒沙门氏菌、副溶血性弧菌)的鉴定。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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