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Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage

BACKGROUND: Polymerase chain reaction (PCR) is used in nucleic acid tests of infectious diseases in point-of-care testing. Previous studies have demonstrated real-time PCR that uses a micro-PCR chip made of packing tape, double-sided tape, and a plastic cover with polycarbonate or polypropylene on a...

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Autores principales: Hwang, Ji-Soo, Kim, Jong-Dae, Kim, Yu-Seop, Song, Hye-Jeong, Park, Chan-Young
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6218989/
https://www.ncbi.nlm.nih.gov/pubmed/30396352
http://dx.doi.org/10.1186/s12938-018-0579-0
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author Hwang, Ji-Soo
Kim, Jong-Dae
Kim, Yu-Seop
Song, Hye-Jeong
Park, Chan-Young
author_facet Hwang, Ji-Soo
Kim, Jong-Dae
Kim, Yu-Seop
Song, Hye-Jeong
Park, Chan-Young
author_sort Hwang, Ji-Soo
collection PubMed
description BACKGROUND: Polymerase chain reaction (PCR) is used in nucleic acid tests of infectious diseases in point-of-care testing. Previous studies have demonstrated real-time PCR that uses a micro-PCR chip made of packing tape, double-sided tape, and a plastic cover with polycarbonate or polypropylene on a black matte printed circuit board substrate. Despite the success of DNA amplification and fluorescence detection using an early version of the micro-PCR chip, reaching the target temperature was fairly slow and, as a result, the total running time was getting longer. To reduce this runtime, the micro-PCR chip was modified by reducing the heater pattern size of the PCB substrate to one-quarter of the original size or less, while maintaining the ability of the heating pattern to cover the reservoir area of the microfluidic channel. In subsequent experiments, DNA amplification failed several times. During the analysis of the cause of this failure, it was found that the reagent was boiling with the heating range from 25 to 95 °C. METHODS: As a method of DNA amplification verification, images were captured by digital single-lens reflex camera to detect FAM fluorescence using diagonal illumination from a blue LED light source. The images were automatically captured at 72 °C (the extension step in nucleic acid amplification) and the brightness of the captured images was analyzed to con-firm the success of DNA amplification. RESULTS: Compared to the previous chip with a larger heating pattern size, the current chip appears to generate excess energy as the size of the heating pattern was reduced. To reduce this excess energy, the initial voltage was lowered to 2 V and 2.5 V, which is equivalent to a one-fifth and one-quarter voltage–power reduction in pulse width modulation control, respectively. In both voltage reduction cases, the DNA amplification was successful. CONCLUSIONS: DNA amplification tests may fail due to the excess energy generated by reducing the heater pattern size of the PCB substrate. However, the tests succeeded when the voltage was reduced to 2 V or 2.5 V. The 2.5 V power test was more efficient for reducing the overall running time.
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spelling pubmed-62189892018-11-08 Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage Hwang, Ji-Soo Kim, Jong-Dae Kim, Yu-Seop Song, Hye-Jeong Park, Chan-Young Biomed Eng Online Research BACKGROUND: Polymerase chain reaction (PCR) is used in nucleic acid tests of infectious diseases in point-of-care testing. Previous studies have demonstrated real-time PCR that uses a micro-PCR chip made of packing tape, double-sided tape, and a plastic cover with polycarbonate or polypropylene on a black matte printed circuit board substrate. Despite the success of DNA amplification and fluorescence detection using an early version of the micro-PCR chip, reaching the target temperature was fairly slow and, as a result, the total running time was getting longer. To reduce this runtime, the micro-PCR chip was modified by reducing the heater pattern size of the PCB substrate to one-quarter of the original size or less, while maintaining the ability of the heating pattern to cover the reservoir area of the microfluidic channel. In subsequent experiments, DNA amplification failed several times. During the analysis of the cause of this failure, it was found that the reagent was boiling with the heating range from 25 to 95 °C. METHODS: As a method of DNA amplification verification, images were captured by digital single-lens reflex camera to detect FAM fluorescence using diagonal illumination from a blue LED light source. The images were automatically captured at 72 °C (the extension step in nucleic acid amplification) and the brightness of the captured images was analyzed to con-firm the success of DNA amplification. RESULTS: Compared to the previous chip with a larger heating pattern size, the current chip appears to generate excess energy as the size of the heating pattern was reduced. To reduce this excess energy, the initial voltage was lowered to 2 V and 2.5 V, which is equivalent to a one-fifth and one-quarter voltage–power reduction in pulse width modulation control, respectively. In both voltage reduction cases, the DNA amplification was successful. CONCLUSIONS: DNA amplification tests may fail due to the excess energy generated by reducing the heater pattern size of the PCB substrate. However, the tests succeeded when the voltage was reduced to 2 V or 2.5 V. The 2.5 V power test was more efficient for reducing the overall running time. BioMed Central 2018-11-06 /pmc/articles/PMC6218989/ /pubmed/30396352 http://dx.doi.org/10.1186/s12938-018-0579-0 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Hwang, Ji-Soo
Kim, Jong-Dae
Kim, Yu-Seop
Song, Hye-Jeong
Park, Chan-Young
Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage
title Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage
title_full Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage
title_fullStr Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage
title_full_unstemmed Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage
title_short Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage
title_sort performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6218989/
https://www.ncbi.nlm.nih.gov/pubmed/30396352
http://dx.doi.org/10.1186/s12938-018-0579-0
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