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Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay
BACKGROUND: A novel avian influenza A (H7N9) virus emerged in eastern China in February 2013. 413 confirmed human cases, including 157 deaths, have been recorded as of July 31, 2014. METHODS: Clinical specimens, including throat swabs, sputum or tracheal aspirates, etc., were obtained from patients...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4286936/ https://www.ncbi.nlm.nih.gov/pubmed/25298249 http://dx.doi.org/10.1186/1471-2334-14-541 |
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author | Fan, Jian Cui, David Lau, Siuying Xie, Guoliang Guo, Xichao Zheng, Shufa Huang, Xiaofeng Yang, Shigui Yang, Xianzhi Huo, Zhaoxia Yu, Fei Lou, Jianzhou Tian, Li Li, Xuefen Dong, Yuejiao Zhu, Qiaoyun Chen, Yu |
author_facet | Fan, Jian Cui, David Lau, Siuying Xie, Guoliang Guo, Xichao Zheng, Shufa Huang, Xiaofeng Yang, Shigui Yang, Xianzhi Huo, Zhaoxia Yu, Fei Lou, Jianzhou Tian, Li Li, Xuefen Dong, Yuejiao Zhu, Qiaoyun Chen, Yu |
author_sort | Fan, Jian |
collection | PubMed |
description | BACKGROUND: A novel avian influenza A (H7N9) virus emerged in eastern China in February 2013. 413 confirmed human cases, including 157 deaths, have been recorded as of July 31, 2014. METHODS: Clinical specimens, including throat swabs, sputum or tracheal aspirates, etc., were obtained from patients exhibiting influenza-like illness (ILIs), especially from those having pneumonia and a history of occupational exposure to poultry and wild birds. RNA was extracted from these samples and a multiplex one-step real-time RT-PCR assay was developed to specifically detect the influenza A virus (FluA). PCR primers targeted the conserved M and Rnase P (RP) genes, as well as the hemagglutinin and neuraminidase genes of the H7N9 virus. RESULTS: The multiplex assay specifically detected the avian H7N9 virus, and no cross-reaction with other common respiratory pathogens was observed. The detection limit of the assay was approximately 0.05 50% tissue culture infective doses (TCID(50)), or 100 copies per reaction. Positive detection of the H7N9 virus in sputum/tracheal aspirates was higher than in throat swabs during the surveillance of patients with ILIs. Additionally, detection of the matrix (M) and Rnase P genes aided in the determination of the novel avian H7N9 virus and ensured the quality of the clinical samples. CONCLUSIONS: These results demonstrate that the multiplex assay detected the novel avian H7N9 virus with high specificity and sensitivity, which is essential for the early diagnosis and treatment of infected patients. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2334-14-541) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4286936 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-42869362015-01-09 Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay Fan, Jian Cui, David Lau, Siuying Xie, Guoliang Guo, Xichao Zheng, Shufa Huang, Xiaofeng Yang, Shigui Yang, Xianzhi Huo, Zhaoxia Yu, Fei Lou, Jianzhou Tian, Li Li, Xuefen Dong, Yuejiao Zhu, Qiaoyun Chen, Yu BMC Infect Dis Research Article BACKGROUND: A novel avian influenza A (H7N9) virus emerged in eastern China in February 2013. 413 confirmed human cases, including 157 deaths, have been recorded as of July 31, 2014. METHODS: Clinical specimens, including throat swabs, sputum or tracheal aspirates, etc., were obtained from patients exhibiting influenza-like illness (ILIs), especially from those having pneumonia and a history of occupational exposure to poultry and wild birds. RNA was extracted from these samples and a multiplex one-step real-time RT-PCR assay was developed to specifically detect the influenza A virus (FluA). PCR primers targeted the conserved M and Rnase P (RP) genes, as well as the hemagglutinin and neuraminidase genes of the H7N9 virus. RESULTS: The multiplex assay specifically detected the avian H7N9 virus, and no cross-reaction with other common respiratory pathogens was observed. The detection limit of the assay was approximately 0.05 50% tissue culture infective doses (TCID(50)), or 100 copies per reaction. Positive detection of the H7N9 virus in sputum/tracheal aspirates was higher than in throat swabs during the surveillance of patients with ILIs. Additionally, detection of the matrix (M) and Rnase P genes aided in the determination of the novel avian H7N9 virus and ensured the quality of the clinical samples. CONCLUSIONS: These results demonstrate that the multiplex assay detected the novel avian H7N9 virus with high specificity and sensitivity, which is essential for the early diagnosis and treatment of infected patients. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2334-14-541) contains supplementary material, which is available to authorized users. BioMed Central 2014-10-08 /pmc/articles/PMC4286936/ /pubmed/25298249 http://dx.doi.org/10.1186/1471-2334-14-541 Text en © Fan et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Fan, Jian Cui, David Lau, Siuying Xie, Guoliang Guo, Xichao Zheng, Shufa Huang, Xiaofeng Yang, Shigui Yang, Xianzhi Huo, Zhaoxia Yu, Fei Lou, Jianzhou Tian, Li Li, Xuefen Dong, Yuejiao Zhu, Qiaoyun Chen, Yu Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay |
title | Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay |
title_full | Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay |
title_fullStr | Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay |
title_full_unstemmed | Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay |
title_short | Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay |
title_sort | detection of a novel avian influenza a (h7n9) virus in humans by multiplex one-step real-time rt-pcr assay |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4286936/ https://www.ncbi.nlm.nih.gov/pubmed/25298249 http://dx.doi.org/10.1186/1471-2334-14-541 |
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