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Early diagnosis of SARS Coronavirus infection by real time RT-PCR
Background: A novel coronavirus was recently identified as the aetiological agent of Severe Acute Respiratory Syndrome (SARS). Molecular assays currently available for detection of SARS-coronavirus (SARS-Cov) have low sensitivity during the early stage of the illness. Objective: To develop and evalu...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Published by Elsevier B.V.
2003
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7129783/ https://www.ncbi.nlm.nih.gov/pubmed/14522060 http://dx.doi.org/10.1016/j.jcv.2003.08.004 |
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author | Poon, Leo L.M. Chan, Kwok Hung Wong, On Kei Yam, Wing Cheong Yuen, Kwok Yung Guan, Yi Lo, Y.M.Dennis Peiris, Joseph S.M. |
author_facet | Poon, Leo L.M. Chan, Kwok Hung Wong, On Kei Yam, Wing Cheong Yuen, Kwok Yung Guan, Yi Lo, Y.M.Dennis Peiris, Joseph S.M. |
author_sort | Poon, Leo L.M. |
collection | PubMed |
description | Background: A novel coronavirus was recently identified as the aetiological agent of Severe Acute Respiratory Syndrome (SARS). Molecular assays currently available for detection of SARS-coronavirus (SARS-Cov) have low sensitivity during the early stage of the illness. Objective: To develop and evaluate a sensitive diagnostic test for SARS by optimizing the viral RNA extraction methods and by applying real-time quantitative RT-PCR technology. Study design: 50 nasopharyngeal aspirate (NPA) samples collected from days 1–3 of disease onset from SARS patients in whom SARS CoV infections was subsequently serologically confirmed and 30 negative control samples were studied. Samples were tested by: (1) our first generation conventional RT-PCR assay with a routine RNA extraction method (Lancet 361 (2003) 1319), (2) our first generation conventional RT-PCR assay with a modified RNA extraction method, (3) a real-time quantitative RT-PCR assay with a modified RNA extraction method. Results: Of 50 NPA specimens collected during the first 3 days of illness, 11 (22%) were positive in our first generation RT-PCR assay. With a modification in the RNA extraction protocol, 22 (44%) samples were positive in the conventional RT-PCR assay. By combining the modified RNA extraction method and real-time quantitative PCR technology, 40 (80%) of these samples were positive in the real-time RT-PCR assay. No positive signal was observed in the negative controls. Conclusion: By optimizing RNA extraction methods and applying quantitative real time RT-PCR technologies, the sensitivity of tests for early diagnosis of SARS can be greatly enhanced. |
format | Online Article Text |
id | pubmed-7129783 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2003 |
publisher | Published by Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71297832020-04-08 Early diagnosis of SARS Coronavirus infection by real time RT-PCR Poon, Leo L.M. Chan, Kwok Hung Wong, On Kei Yam, Wing Cheong Yuen, Kwok Yung Guan, Yi Lo, Y.M.Dennis Peiris, Joseph S.M. J Clin Virol Article Background: A novel coronavirus was recently identified as the aetiological agent of Severe Acute Respiratory Syndrome (SARS). Molecular assays currently available for detection of SARS-coronavirus (SARS-Cov) have low sensitivity during the early stage of the illness. Objective: To develop and evaluate a sensitive diagnostic test for SARS by optimizing the viral RNA extraction methods and by applying real-time quantitative RT-PCR technology. Study design: 50 nasopharyngeal aspirate (NPA) samples collected from days 1–3 of disease onset from SARS patients in whom SARS CoV infections was subsequently serologically confirmed and 30 negative control samples were studied. Samples were tested by: (1) our first generation conventional RT-PCR assay with a routine RNA extraction method (Lancet 361 (2003) 1319), (2) our first generation conventional RT-PCR assay with a modified RNA extraction method, (3) a real-time quantitative RT-PCR assay with a modified RNA extraction method. Results: Of 50 NPA specimens collected during the first 3 days of illness, 11 (22%) were positive in our first generation RT-PCR assay. With a modification in the RNA extraction protocol, 22 (44%) samples were positive in the conventional RT-PCR assay. By combining the modified RNA extraction method and real-time quantitative PCR technology, 40 (80%) of these samples were positive in the real-time RT-PCR assay. No positive signal was observed in the negative controls. Conclusion: By optimizing RNA extraction methods and applying quantitative real time RT-PCR technologies, the sensitivity of tests for early diagnosis of SARS can be greatly enhanced. Published by Elsevier B.V. 2003-12 2003-09-24 /pmc/articles/PMC7129783/ /pubmed/14522060 http://dx.doi.org/10.1016/j.jcv.2003.08.004 Text en Copyright © 2003 Published by Elsevier B.V. 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 Poon, Leo L.M. Chan, Kwok Hung Wong, On Kei Yam, Wing Cheong Yuen, Kwok Yung Guan, Yi Lo, Y.M.Dennis Peiris, Joseph S.M. Early diagnosis of SARS Coronavirus infection by real time RT-PCR |
title | Early diagnosis of SARS Coronavirus infection by real time RT-PCR |
title_full | Early diagnosis of SARS Coronavirus infection by real time RT-PCR |
title_fullStr | Early diagnosis of SARS Coronavirus infection by real time RT-PCR |
title_full_unstemmed | Early diagnosis of SARS Coronavirus infection by real time RT-PCR |
title_short | Early diagnosis of SARS Coronavirus infection by real time RT-PCR |
title_sort | early diagnosis of sars coronavirus infection by real time rt-pcr |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7129783/ https://www.ncbi.nlm.nih.gov/pubmed/14522060 http://dx.doi.org/10.1016/j.jcv.2003.08.004 |
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