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SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses
Coronaviruses are etiologic agents of respiratory and enteric diseases in humans and in animals. In this study, a one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on SYBR Green chemistry and degenerate primers was developed for the generic detection of coronavi...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer-Verlag
2006
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087200/ https://www.ncbi.nlm.nih.gov/pubmed/16941059 http://dx.doi.org/10.1007/s00705-006-0840-x |
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author | Escutenaire, S. Mohamed, N. Isaksson, M. Thorén, P. Klingeborn, B. Belák, S. Berg, M. Blomberg, J. |
author_facet | Escutenaire, S. Mohamed, N. Isaksson, M. Thorén, P. Klingeborn, B. Belák, S. Berg, M. Blomberg, J. |
author_sort | Escutenaire, S. |
collection | PubMed |
description | Coronaviruses are etiologic agents of respiratory and enteric diseases in humans and in animals. In this study, a one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on SYBR Green chemistry and degenerate primers was developed for the generic detection of coronaviruses. The primers, designed in the open reading frame 1b, enabled the detection of 32 animal coronaviruses including strains of canine coronavirus, feline coronavirus, transmissible gastroenteritis virus (TGEV), bovine coronavirus (BCoV), murine hepatitis virus (MHV) and infectious bronchitis virus (IBV). A specific amplification was also observed with the human coronaviruses (HCoV) HCoV-NL63, HCoV-OC43, HCoV-229E and severe acute respiratory syndrome coronavirus (SARS-CoV). The real-time RT-PCR detected down to 10 cRNA copies from TGEV, BCoV, SARS-CoV and IBV. In addition, the assay exhibited a high sensitivity and specificity on clinical samples from different animal species. The developed assay represents a potential tool for laboratory diagnostics and for detecting still uncharacterized coronaviruses. |
format | Online Article Text |
id | pubmed-7087200 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | Springer-Verlag |
record_format | MEDLINE/PubMed |
spelling | pubmed-70872002020-03-23 SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses Escutenaire, S. Mohamed, N. Isaksson, M. Thorén, P. Klingeborn, B. Belák, S. Berg, M. Blomberg, J. Arch Virol Article Coronaviruses are etiologic agents of respiratory and enteric diseases in humans and in animals. In this study, a one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on SYBR Green chemistry and degenerate primers was developed for the generic detection of coronaviruses. The primers, designed in the open reading frame 1b, enabled the detection of 32 animal coronaviruses including strains of canine coronavirus, feline coronavirus, transmissible gastroenteritis virus (TGEV), bovine coronavirus (BCoV), murine hepatitis virus (MHV) and infectious bronchitis virus (IBV). A specific amplification was also observed with the human coronaviruses (HCoV) HCoV-NL63, HCoV-OC43, HCoV-229E and severe acute respiratory syndrome coronavirus (SARS-CoV). The real-time RT-PCR detected down to 10 cRNA copies from TGEV, BCoV, SARS-CoV and IBV. In addition, the assay exhibited a high sensitivity and specificity on clinical samples from different animal species. The developed assay represents a potential tool for laboratory diagnostics and for detecting still uncharacterized coronaviruses. Springer-Verlag 2006-08-28 2007 /pmc/articles/PMC7087200/ /pubmed/16941059 http://dx.doi.org/10.1007/s00705-006-0840-x Text en © Springer-Verlag 2006 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Article Escutenaire, S. Mohamed, N. Isaksson, M. Thorén, P. Klingeborn, B. Belák, S. Berg, M. Blomberg, J. SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses |
title | SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses |
title_full | SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses |
title_fullStr | SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses |
title_full_unstemmed | SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses |
title_short | SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses |
title_sort | sybr green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087200/ https://www.ncbi.nlm.nih.gov/pubmed/16941059 http://dx.doi.org/10.1007/s00705-006-0840-x |
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