3D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recovery

Molecular diagnosis is a gold standard method for identifying an infectious disease. DNA extraction from a target pathogen is one of the most important procedures for accurate analysis of the disease-causative pathogen. In this study, a novel 3D-printed electrophoretic DNA extraction microdevice (3D...

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Main Authors: Kiwon Nam, Seungbeom Kim, Younseong Song, Yoo Seok Lee, Seok Jae Lee, Kyoung G. Lee, Yong Tae Kim
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Sensors and Actuators Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666053924000602
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author Kiwon Nam
Seungbeom Kim
Younseong Song
Yoo Seok Lee
Seok Jae Lee
Kyoung G. Lee
Yong Tae Kim
author_facet Kiwon Nam
Seungbeom Kim
Younseong Song
Yoo Seok Lee
Seok Jae Lee
Kyoung G. Lee
Yong Tae Kim
author_sort Kiwon Nam
collection DOAJ
description Molecular diagnosis is a gold standard method for identifying an infectious disease. DNA extraction from a target pathogen is one of the most important procedures for accurate analysis of the disease-causative pathogen. In this study, a novel 3D-printed electrophoretic DNA extraction microdevice (3D-EDEM) was developed using a digital light processing-stereolithography (DLP-SL) for point-of-care analysis. The 3D-EDEM consists of a source chamber for a bacteria lysate reservoir, a sink chamber for an elution solution container, a hydrogel channel embracing capillary channels that act as a sieving matrix for size-based separation, and two electrode holders for supplying electrical current. Prior to fabricating the 3D-EDEM, UV-curable resin was prepared by using a poly(ethylene glycol) diacrylate (PEG-DA), Irgarcure 819 (IRG), and 2-isopropyl thioxanthone (ITX) as a monomer, a photoinitiator, and a photosensitizer, respectively. The 3D-printed 3D-EDEM provides numerous merits of being inexpensive, reproducible, and convenient, making it more suitable for on-site DNA extraction microdevices than soft-lithographic procedures. For DNA extraction on the 3D-EDEM, Escherichia coli O157:H7 (E. coli) lysate and elution buffer were loaded into the source chamber and the sink chamber, respectively. The optimum DNA extraction time and limit of the DNA extraction test of 3D-EDEM were carried out to evaluate DNA extraction performance, especially using a portable battery. Additionally, the successful DNA extraction test from artificially infected food samples confirms the applicability of the 3D-EDEM to real fields. The proposed 3D-EDEM is adequate for on-site DNA extraction in the field of clinical diagnosis, food safety, environmental monitoring, and forensic analysis.
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spelling doaj-art-d60cb60ecb1840dda842749c3a54e13d2024-11-15T06:15:04ZengElsevierSensors and Actuators Reports2666-05392024-12-0181002443D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recoveryKiwon Nam0Seungbeom Kim1Younseong Song2Yoo Seok Lee3Seok Jae Lee4Kyoung G. Lee5Yong Tae Kim6Department of Chemical Engineering & Biotechnology, Tech University of Korea, 237 Sangidaehak-ro, Siheung-si, Gyeonggi-do 15073, Republic of KoreaDepartment of Chemical Engineering & Biotechnology, Tech University of Korea, 237 Sangidaehak-ro, Siheung-si, Gyeonggi-do 15073, Republic of KoreaNanobio Application Team, National Nanofab Center, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of KoreaDepartment of Chemical Engineering & Biotechnology, Tech University of Korea, 237 Sangidaehak-ro, Siheung-si, Gyeonggi-do 15073, Republic of KoreaNanobio Application Team, National Nanofab Center, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of KoreaNanobio Application Team, National Nanofab Center, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea; Corresponding authors.Department of Chemical Engineering & Biotechnology, Tech University of Korea, 237 Sangidaehak-ro, Siheung-si, Gyeonggi-do 15073, Republic of Korea; Corresponding authors.Molecular diagnosis is a gold standard method for identifying an infectious disease. DNA extraction from a target pathogen is one of the most important procedures for accurate analysis of the disease-causative pathogen. In this study, a novel 3D-printed electrophoretic DNA extraction microdevice (3D-EDEM) was developed using a digital light processing-stereolithography (DLP-SL) for point-of-care analysis. The 3D-EDEM consists of a source chamber for a bacteria lysate reservoir, a sink chamber for an elution solution container, a hydrogel channel embracing capillary channels that act as a sieving matrix for size-based separation, and two electrode holders for supplying electrical current. Prior to fabricating the 3D-EDEM, UV-curable resin was prepared by using a poly(ethylene glycol) diacrylate (PEG-DA), Irgarcure 819 (IRG), and 2-isopropyl thioxanthone (ITX) as a monomer, a photoinitiator, and a photosensitizer, respectively. The 3D-printed 3D-EDEM provides numerous merits of being inexpensive, reproducible, and convenient, making it more suitable for on-site DNA extraction microdevices than soft-lithographic procedures. For DNA extraction on the 3D-EDEM, Escherichia coli O157:H7 (E. coli) lysate and elution buffer were loaded into the source chamber and the sink chamber, respectively. The optimum DNA extraction time and limit of the DNA extraction test of 3D-EDEM were carried out to evaluate DNA extraction performance, especially using a portable battery. Additionally, the successful DNA extraction test from artificially infected food samples confirms the applicability of the 3D-EDEM to real fields. The proposed 3D-EDEM is adequate for on-site DNA extraction in the field of clinical diagnosis, food safety, environmental monitoring, and forensic analysis.http://www.sciencedirect.com/science/article/pii/S26660539240006023D-printingDNA extractionElectrophoreticMicrofluidicsGenetic analysis
spellingShingle Kiwon Nam
Seungbeom Kim
Younseong Song
Yoo Seok Lee
Seok Jae Lee
Kyoung G. Lee
Yong Tae Kim
3D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recovery
Sensors and Actuators Reports
3D-printing
DNA extraction
Electrophoretic
Microfluidics
Genetic analysis
title 3D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recovery
title_full 3D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recovery
title_fullStr 3D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recovery
title_full_unstemmed 3D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recovery
title_short 3D-printable electrophoretic DNA extraction microdevice for on-site bacterial DNA recovery
title_sort 3d printable electrophoretic dna extraction microdevice for on site bacterial dna recovery
topic 3D-printing
DNA extraction
Electrophoretic
Microfluidics
Genetic analysis
url http://www.sciencedirect.com/science/article/pii/S2666053924000602
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