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用于即时检测多种食源性致病菌的全集成可滑动纸嵌入塑料微器件。

Fully integrated and slidable paper-embedded plastic microdevice for point-of-care testing of multiple foodborne pathogens.

机构信息

Department of BioNano Technology, Gachon University 1342 Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do, 13120, South Korea.

Department of BioNano Technology, Gachon University 1342 Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do, 13120, South Korea.

出版信息

Biosens Bioelectron. 2019 Jun 15;135:120-128. doi: 10.1016/j.bios.2019.04.011. Epub 2019 Apr 11.

Abstract

This study presents a slidable paper-embedded plastic microdevice fully integrated with DNA extraction, loop-mediated isothermal amplification (LAMP), and colorimetric detection functionalities. The developed microdevice consists of three layers that allow a sliding movement to mix the sample and reagents for DNA purification, amplification, and detection in a sequential manner. An FTA card was employed in the main chamber for DNA extraction and purification from intact bacterial cells. Subsequently, LAMP reagents and fuchsin-stored chambers were pulled toward the main chambers for DNA amplifications at 65 °C. After 30 min, the detection reagents-stored chambers were then moved to main chambers for result analysis. For the detection of LAMP amplicons, a novel colorimetric fuchsin-based method was employed. The wide applicability of the integrated microdevice was demonstrated by successfully screening three major foodborne pathogens, namely Salmonella spp., Staphylococcus aureus, and Escherichia coli O157:H7 in food, enabling highly sensitive detection of 3.0 × 10 CFU/sample of Gram-negative bacteria (Salmonella spp. and Escherichia coli O157:H7) and 3.0 × 10 CFU/sample of Gram-positive bacteria (Staphylococcus aureus) within 75 min. The portable and integrated microdevice presented in this study holds significant promise for point-of-care applications to accurately and rapidly diagnose and control diseases.

摘要

本研究提出了一种可滑动的纸质嵌入式塑料微器件,该微器件完全集成了 DNA 提取、环介导等温扩增(LAMP)和比色检测功能。所开发的微器件由三层组成,允许滑动运动以顺序混合样品和试剂,用于 DNA 纯化、扩增和检测。FTA 卡被用于主腔室中,用于从完整的细菌细胞中提取和纯化 DNA。随后,将 LAMP 试剂和藏红花试剂腔室拉向主腔室,在 65°C 下进行 DNA 扩增。30 分钟后,检测试剂腔室然后移动到主腔室进行结果分析。对于 LAMP 扩增子的检测,采用了一种新颖的基于比色藏红花的方法。该集成微器件的广泛适用性通过成功筛选三种主要食源性病原体(即沙门氏菌、金黄色葡萄球菌和大肠杆菌 O157:H7)得到了证明,能够高度敏感地检测到 3.0×10 CFU/样品的革兰氏阴性菌(沙门氏菌和大肠杆菌 O157:H7)和 3.0×10 CFU/样品的革兰氏阳性菌(金黄色葡萄球菌),时间在 75 分钟内。本研究中提出的便携式集成微器件有望在即时护理应用中实现准确、快速的疾病诊断和控制。

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