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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...

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Autores principales: Pyrc, Krzysztof, Milewska, Aleksandra, Potempa, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. Published by Elsevier B.V. 2011
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.
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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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