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Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63
Human coronavirus NL63 was identified in 2004 in the Netherlands. Due to the high prevalence and world-wide distribution of this pathogen, it is essential to develop a sensitive and specific detection assay suitable for use in a routine diagnostic laboratory. Techniques based on PCR or real-time PCR...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V. Published by Elsevier B.V.
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112811/ https://www.ncbi.nlm.nih.gov/pubmed/21545810 http://dx.doi.org/10.1016/j.jviromet.2011.04.024 |
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author | Pyrc, Krzysztof Milewska, Aleksandra Potempa, Jan |
author_facet | Pyrc, Krzysztof Milewska, Aleksandra Potempa, Jan |
author_sort | Pyrc, Krzysztof |
collection | PubMed |
description | Human coronavirus NL63 was identified in 2004 in the Netherlands. Due to the high prevalence and world-wide distribution of this pathogen, it is essential to develop a sensitive and specific detection assay suitable for use in a routine diagnostic laboratory. Techniques based on PCR or real-time PCR are laborious and expensive. Detailed analysis of the HCoV-NL63 genome permitted the identification of a conserved nucleic acid sequential motif, which was sufficient for the design of a loop-mediated isothermal amplification (LAMP) assay. Evaluation of the method showed that the test is specific to HCoV-NL63 and that it does not cross-react with other respiratory viruses. The detection limit was found to be 1 copy of RNA template per reaction in cell culture supernatants and clinical specimens. |
format | Online Article Text |
id | pubmed-7112811 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Elsevier B.V. Published by Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71128112020-04-02 Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63 Pyrc, Krzysztof Milewska, Aleksandra Potempa, Jan J Virol Methods Article Human coronavirus NL63 was identified in 2004 in the Netherlands. Due to the high prevalence and world-wide distribution of this pathogen, it is essential to develop a sensitive and specific detection assay suitable for use in a routine diagnostic laboratory. Techniques based on PCR or real-time PCR are laborious and expensive. Detailed analysis of the HCoV-NL63 genome permitted the identification of a conserved nucleic acid sequential motif, which was sufficient for the design of a loop-mediated isothermal amplification (LAMP) assay. Evaluation of the method showed that the test is specific to HCoV-NL63 and that it does not cross-react with other respiratory viruses. The detection limit was found to be 1 copy of RNA template per reaction in cell culture supernatants and clinical specimens. Elsevier B.V. Published by Elsevier B.V. 2011-07 2011-04-27 /pmc/articles/PMC7112811/ /pubmed/21545810 http://dx.doi.org/10.1016/j.jviromet.2011.04.024 Text en Copyright © 2011 Elsevier B.V. Published by Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Pyrc, Krzysztof Milewska, Aleksandra Potempa, Jan Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63 |
title | Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63 |
title_full | Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63 |
title_fullStr | Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63 |
title_full_unstemmed | Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63 |
title_short | Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63 |
title_sort | development of loop-mediated isothermal amplification assay for detection of human coronavirus-nl63 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112811/ https://www.ncbi.nlm.nih.gov/pubmed/21545810 http://dx.doi.org/10.1016/j.jviromet.2011.04.024 |
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