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Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection

We developed an integrated PCR system that performs automated sample preparation and fast polymerase chain reaction (PCR) for application in point-of care (POC) testing. This system is assembled from inexpensive 3D-printing parts, off-the-shelf electronics and motors. Molecular detection requires a...

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Autores principales: Lee, Dongkyu, Kim, Deawook, Han, Jounghyuk, Yun, Jongsu, Lee, Kang-Ho, Kim, Gyu Man, Kwon, Ohwon, Lee, Jaejong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7827619/
https://www.ncbi.nlm.nih.gov/pubmed/33430443
http://dx.doi.org/10.3390/s21020377
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author Lee, Dongkyu
Kim, Deawook
Han, Jounghyuk
Yun, Jongsu
Lee, Kang-Ho
Kim, Gyu Man
Kwon, Ohwon
Lee, Jaejong
author_facet Lee, Dongkyu
Kim, Deawook
Han, Jounghyuk
Yun, Jongsu
Lee, Kang-Ho
Kim, Gyu Man
Kwon, Ohwon
Lee, Jaejong
author_sort Lee, Dongkyu
collection PubMed
description We developed an integrated PCR system that performs automated sample preparation and fast polymerase chain reaction (PCR) for application in point-of care (POC) testing. This system is assembled from inexpensive 3D-printing parts, off-the-shelf electronics and motors. Molecular detection requires a series of procedures including sample preparation, amplification, and fluorescence intensity analysis. The system can perform automated DNA sample preparation (extraction, separation and purification) in ≤5 min. The variance of the automated sample preparation was clearly lower than that achieved using manual DNA extraction. Fast thermal ramp cycles were generated by a customized thermocycler designed to automatically transport samples between heating and cooling blocks. Despite the large sample volume (50 μL), rapid two-step PCR amplification completed 40 cycles in ≤13.8 min. Variations in fluorescence intensity were measured by analyzing fluorescence images. As proof of concept of this system, we demonstrated the rapid DNA detection of pathogenic bacteria. We also compared the sensitivity of this system with that of a commercial device during the automated extraction and fast PCR of Salmonella bacteria.
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spelling pubmed-78276192021-01-25 Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection Lee, Dongkyu Kim, Deawook Han, Jounghyuk Yun, Jongsu Lee, Kang-Ho Kim, Gyu Man Kwon, Ohwon Lee, Jaejong Sensors (Basel) Letter We developed an integrated PCR system that performs automated sample preparation and fast polymerase chain reaction (PCR) for application in point-of care (POC) testing. This system is assembled from inexpensive 3D-printing parts, off-the-shelf electronics and motors. Molecular detection requires a series of procedures including sample preparation, amplification, and fluorescence intensity analysis. The system can perform automated DNA sample preparation (extraction, separation and purification) in ≤5 min. The variance of the automated sample preparation was clearly lower than that achieved using manual DNA extraction. Fast thermal ramp cycles were generated by a customized thermocycler designed to automatically transport samples between heating and cooling blocks. Despite the large sample volume (50 μL), rapid two-step PCR amplification completed 40 cycles in ≤13.8 min. Variations in fluorescence intensity were measured by analyzing fluorescence images. As proof of concept of this system, we demonstrated the rapid DNA detection of pathogenic bacteria. We also compared the sensitivity of this system with that of a commercial device during the automated extraction and fast PCR of Salmonella bacteria. MDPI 2021-01-07 /pmc/articles/PMC7827619/ /pubmed/33430443 http://dx.doi.org/10.3390/s21020377 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Letter
Lee, Dongkyu
Kim, Deawook
Han, Jounghyuk
Yun, Jongsu
Lee, Kang-Ho
Kim, Gyu Man
Kwon, Ohwon
Lee, Jaejong
Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection
title Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection
title_full Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection
title_fullStr Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection
title_full_unstemmed Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection
title_short Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection
title_sort integrated, automated, fast pcr system for point-of-care molecular diagnosis of bacterial infection
topic Letter
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7827619/
https://www.ncbi.nlm.nih.gov/pubmed/33430443
http://dx.doi.org/10.3390/s21020377
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