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Issue 8, 2020
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An electrophoretic DNA extraction device using a nanofilter for molecular diagnosis of pathogens

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Abstract

Rapid and efficient nucleic acid (NA) extraction and concentration are required for point-of-care analysis in order to prevent an epidemic/pandemic disease outbreak. Typical silica-based NA extraction methods have limitations such as being time-consuming, requiring human intervention, and resulting in a low recovery yield. In this study, we have developed a pathogenic DNA extraction device based on electrokinetic separation incorporated with a silicon nitride (SiNx) nanofilter, which expedites the DNA extraction procedure with advantages of being convenient, efficient, and inexpensive. This DNA extraction device consists of a computer numerical control (CNC) milled-Teflon gadget with a cis-chamber as a cell lysate reservoir and a trans-chamber as a elution solution reservoir, with the SiNx nanofilter being inserted between the two chambers. The SiNx nanofilter was fabricated using a photolithographic method in conjunction with nanoimprinting. Approximately 7.2 million nanopores of 220 nm diameter were located at the center of the nanofilter. When a DC electric field is applied through the nanopores, DNA is transferred from the cis-chamber to the trans-chamber to isolate the DNA from the cell debris. To demonstrate the DNA extraction performance, we measured the absorbances at 260 and 280 nm and performed a real-time polymerase chain reaction (real-time PCR) using the recovered DNA to verify its feasibility for downstream genetic analysis. Moreover, the DNA extraction device was successfully operated using a 1.5 V alkaline battery, which verifies the portability of the device for point-of-care testing. Such an advanced DNA extraction system can be utilized in various fields including clinical analysis, pathogen detection, forensic analysis, and on-site detection.

Graphical abstract: An electrophoretic DNA extraction device using a nanofilter for molecular diagnosis of pathogens

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Supplementary files

Article information


Submitted
18 Dec 2019
Accepted
07 Feb 2020
First published
10 Feb 2020

Nanoscale, 2020,12, 5048-5054
Article type
Paper

An electrophoretic DNA extraction device using a nanofilter for molecular diagnosis of pathogens

J. Kang, Y. T. Kim, K. Lee, H. Kim, K. G. Lee, J. Ahn, J. Lee, S. J. Lee and K. Kim, Nanoscale, 2020, 12, 5048
DOI: 10.1039/C9NR10675H

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