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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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. |
format | Online Article Text |
id | pubmed-7827619 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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