Disposable, pressure-driven, and self-contained cartridge with pre-stored reagents for automated nucleic acid extraction
Artikel i vetenskaplig tidskrift, 2023

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.

Bacteria

Pathogen detection

Extraction

Purification

Cartridge

Nucleic acid

Författare

Le Tran Huy Thang

Pukyong National University

Won Han

Pukyong National University

Jae Ho Shin

Chalmers, Life sciences, Industriell bioteknik

Joong Ho Shin

Pukyong National University

Sensors and Actuators, B: Chemical

0925-4005 (ISSN)

Vol. 375 132948

Ämneskategorier

Analytisk kemi

Annan medicinsk grundvetenskap

Annan industriell bioteknik

DOI

10.1016/j.snb.2022.132948

Mer information

Senast uppdaterat

2022-11-25