Disposable, pressure-driven, and self-contained cartridge with pre-stored reagents for automated nucleic acid extraction

墨盒 核酸法 微流控 核酸 萃取(化学) 溶解 核酸定量 色谱法 DNA提取 试剂 固相萃取 化学 DNA 样品制备 聚合酶链反应 纳米技术 生物化学 材料科学 基因 冶金 物理化学
作者
Le Tran Huy Thang,Won Seok Han,Jaeho Shin,Joong Ho Shin
出处
期刊:Sensors and Actuators B-chemical [Elsevier BV]
卷期号:375: 132948-132948 被引量:6
标识
DOI:10.1016/j.snb.2022.132948
摘要

Nucleic acid extraction is vital in many applications such as molecular diagnostics, genetic engineering, and deoxyribonucleic acid (DNA) sequencing. Although recent advances in nucleic acid extraction have been made to improve the process using microfluidics, it is still necessary to make it easy for end-users by having pre-loaded reagents, eliminating pipetting between steps, and automating the process for point-of-care (POC) testing. Herein, we present a pressure-driven and self-contained cartridge with pre-stored reagents for automating nucleic acid purification. To reduce operational complexity, reagents were transferred through a microfluidic chip using pressurized air stored inside the cartridge instead of an external pump or valving system. After performing cell lysis, the cartridge was inserted into the device, and the nucleic acid was purified automatically within 3 min Escherichia coli (E. coli) O157:H7 DNA extracted by our device showed similar concentration, purity, and real-time polymerase chain reaction (qPCR) results as the conventional column-based nucleic acid extraction method. Our device achieved a detection limit of 103 CFU for E. coli DNA, which is the same as that obtained using the conventional solid-phase extraction method. This study introduces a novel approach for automating the sample preparation process, which can help in facilitating POC molecular diagnostics.
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