Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction
The current report devised a novel isothermal diagnostic assay, termed as nanoparticle-based biosensor (NB)- and antarctic thermal sensitive uracil-DNA-glycosylase (ATSU)-supplemented polymerase spiral reaction (PSR; NB-ATSU-PSR). The technique merges enzymatic digestion of carryover contaminants an...
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ftpubmed:oai:pubmedcentral.nih.gov:6923221 2023-05-15T14:02:58+02:00 Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction Wang, Yi Jiao, Wei-wei Wang, Yacui Sun, Lin Li, Jie-qiong Wang, Ze-ming Xiao, Jing Shen, Chen Xu, Fang Qi, Hui Wang, Yong-hong Guo, Ya-jie Shen, A-dong 2019-12-13 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923221/ https://doi.org/10.3389/fbioe.2019.00401 en eng Frontiers Media S.A. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923221/ http://dx.doi.org/10.3389/fbioe.2019.00401 Copyright © 2019 Wang, Jiao, Wang, Sun, Li, Wang, Xiao, Shen, Xu, Qi, Wang, Guo and Shen. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. CC-BY Bioengineering and Biotechnology Text 2019 ftpubmed https://doi.org/10.3389/fbioe.2019.00401 2020-01-12T01:16:28Z The current report devised a novel isothermal diagnostic assay, termed as nanoparticle-based biosensor (NB)- and antarctic thermal sensitive uracil-DNA-glycosylase (ATSU)-supplemented polymerase spiral reaction (PSR; NB-ATSU-PSR). The technique merges enzymatic digestion of carryover contaminants and isothermal nucleic acid amplification technique (PSR) for simultaneous detection of nucleic acid sequences and elimination of carryover contamination. In particular, nucleic acid amplification and elimination of carryover contamination are conducted in a single pot and, thus, the use of a closed-tube reaction can remove undesired results due to carryover contamination. For demonstration purpose, Klebsiella pneumoniae is employed as the model to demonstrate the usability of NB-ATSU-PSR assay. The assay's sensitivity, specificity, and practical feasibility were successfully evaluated using the pure cultures and sputum samples. The amplification products were detectable from as little as 100 fg of genomic DNAs and from ~550 colony-forming unit (CFU) in 1 ml of spiked sputum samples. All K. pneumoniae strains examined were positive for NB-ATSU-PSR detection, and all non-K. pneumoniae strains tested were negative for the NB-ATSU-PSR technique. The whole process, including rapid template preparation (20 min), PSR amplification (60 min), ATSU treatment (5 min), and result reporting (within 2 min), can be finished within 90 min. As a proof-of-concept methodology, NB-ATSU-PSR technique can be reconfigured to detect various target nucleic acid sequences by redesigning the PSR primer set. Text Antarc* Antarctic PubMed Central (PMC) Antarctic Frontiers in Bioengineering and Biotechnology 7 |
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Bioengineering and Biotechnology |
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Bioengineering and Biotechnology Wang, Yi Jiao, Wei-wei Wang, Yacui Sun, Lin Li, Jie-qiong Wang, Ze-ming Xiao, Jing Shen, Chen Xu, Fang Qi, Hui Wang, Yong-hong Guo, Ya-jie Shen, A-dong Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction |
topic_facet |
Bioengineering and Biotechnology |
description |
The current report devised a novel isothermal diagnostic assay, termed as nanoparticle-based biosensor (NB)- and antarctic thermal sensitive uracil-DNA-glycosylase (ATSU)-supplemented polymerase spiral reaction (PSR; NB-ATSU-PSR). The technique merges enzymatic digestion of carryover contaminants and isothermal nucleic acid amplification technique (PSR) for simultaneous detection of nucleic acid sequences and elimination of carryover contamination. In particular, nucleic acid amplification and elimination of carryover contamination are conducted in a single pot and, thus, the use of a closed-tube reaction can remove undesired results due to carryover contamination. For demonstration purpose, Klebsiella pneumoniae is employed as the model to demonstrate the usability of NB-ATSU-PSR assay. The assay's sensitivity, specificity, and practical feasibility were successfully evaluated using the pure cultures and sputum samples. The amplification products were detectable from as little as 100 fg of genomic DNAs and from ~550 colony-forming unit (CFU) in 1 ml of spiked sputum samples. All K. pneumoniae strains examined were positive for NB-ATSU-PSR detection, and all non-K. pneumoniae strains tested were negative for the NB-ATSU-PSR technique. The whole process, including rapid template preparation (20 min), PSR amplification (60 min), ATSU treatment (5 min), and result reporting (within 2 min), can be finished within 90 min. As a proof-of-concept methodology, NB-ATSU-PSR technique can be reconfigured to detect various target nucleic acid sequences by redesigning the PSR primer set. |
format |
Text |
author |
Wang, Yi Jiao, Wei-wei Wang, Yacui Sun, Lin Li, Jie-qiong Wang, Ze-ming Xiao, Jing Shen, Chen Xu, Fang Qi, Hui Wang, Yong-hong Guo, Ya-jie Shen, A-dong |
author_facet |
Wang, Yi Jiao, Wei-wei Wang, Yacui Sun, Lin Li, Jie-qiong Wang, Ze-ming Xiao, Jing Shen, Chen Xu, Fang Qi, Hui Wang, Yong-hong Guo, Ya-jie Shen, A-dong |
author_sort |
Wang, Yi |
title |
Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction |
title_short |
Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction |
title_full |
Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction |
title_fullStr |
Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction |
title_full_unstemmed |
Simultaneous Nucleic Acids Detection and Elimination of Carryover Contamination With Nanoparticles-Based Biosensor- and Antarctic Thermal Sensitive Uracil-DNA-Glycosylase-Supplemented Polymerase Spiral Reaction |
title_sort |
simultaneous nucleic acids detection and elimination of carryover contamination with nanoparticles-based biosensor- and antarctic thermal sensitive uracil-dna-glycosylase-supplemented polymerase spiral reaction |
publisher |
Frontiers Media S.A. |
publishDate |
2019 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923221/ https://doi.org/10.3389/fbioe.2019.00401 |
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Antarctic |
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Antarctic |
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Antarc* Antarctic |
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Antarc* Antarctic |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923221/ http://dx.doi.org/10.3389/fbioe.2019.00401 |
op_rights |
Copyright © 2019 Wang, Jiao, Wang, Sun, Li, Wang, Xiao, Shen, Xu, Qi, Wang, Guo and Shen. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.3389/fbioe.2019.00401 |
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Frontiers in Bioengineering and Biotechnology |
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1766273422666498048 |