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Simultaneous Detection of 2019 Novel Coronavirus and Influenza Virus by Double Fluorescent RT-PCR

Received: 6 March 2020     Accepted: 26 March 2020     Published: 13 April 2020
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Abstract

This paper introduces a method for simultaneous detection of 2019 novel coronavirus (2019-nCoV) and influenza virus A and influenza virus B (FluA/B) by dual fluorescent RT-PCR, providing some references for the current clinical first-line practice against the epidemic. More than 12000 samples of nasopharyngeal swabs, sputum and anal swabs were collected. All samples were inactivated at 60°C for 40 min before testing, so as to protect medical examine. Nucleic acid detection kits from two manufacturers of 2019-nCoV and FluA/B were selected for the detection. Carboxyfluorescein (FAM) and green fluorescent protein (VIC) labeled probes were used to achieve simultaneous detection of the four gene targets using a double fluorescent RT-PCR reaction system. According to the results of nucleic acid detection, there is no cross infection between 2019-nCoV and FluA/B. The CT value of novel coronavirus nucleic acid in anal swab greater than CT value of sputum greater than CT value of nasopharyngeal swab in the same patient. This paper introduces a method for rapid and simultaneous detection of 2019-nCoV and FluA/B by dual fluorescent RT-PCR. It was improved the detection efficiency and reduce the cost, which could be used for rapid and emergent detection of 2019-nCoV and FluA/B. Our experiment is fast and accurate, could make reference for nucleic acid detection medical diagnosis for clinical medical staff.

Published in American Journal of Laboratory Medicine (Volume 5, Issue 1)
DOI 10.11648/j.ajlm.20200501.16
Page(s) 42-46
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2020. Published by Science Publishing Group

Keywords

2019-nCoV, A/B Influenza Virus, Nucleic Acid Detection, Double Fluorescent RT-PCR

References
[1] Zhu N, Zhang D, Wang W, Li X, et al. A Novel Coronavirus from Patients with Pneumonia in China, 2019 N Engl J Med. 2020 pp. 727-733.
[2] Huang C, Wang Y, Li X, et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet. 2020 pp. 497-506.
[3] Dayoo Ocean network https://baijiahao.baidu.com/s?id=1659869159132622751&wfr=spider&for=pc
[4] Zhang N, Wang L, Deng X, et al. Recent advances in the detection of respiratory virus infection in humans. J Med Virol. 2020; 92 pp. 408-417.
[5] General Office of China Health and Health Commission, State Administration of Traditional Chinese Medicine. Technical guidelines for laboratory testing of COVID-19 (second edition) (2020-01-22) http://www.nhc.gov.cn/xcs/zhengcwj/202001/c67cfe29ecf1470e8c7fc47d3b751e88.shtml
[6] General Office of China Health and Health Commission, State Administration of Traditional Chinese Medicine. Laboratory biosafety guidelines of 2019-nCoV (second edition) (2020-01-23) http://www.nhc.gov.cn/qjjys/s7948/202001/0909555408d842a58828611dde2e6a26.shtml
[7] Ministry of Health of the PRC Diagnostic criteria for influenza WS285-2008.
[8] phoenix http://feng.ifeng.com/c/7uPEzrxc9Bi
[9] Beijing daily client http://mbd.baidu.com/newspage/data/landingsuper?context=%7B%22nid%22%3A%22news_8695525602424460969%22%7D&n_type=0&p_from=1
[10] General Office of China Health and Health Commission, State Administration of Traditional Chinese Medicine. Diagnosis and treatment plan of COVID-19 (Trial version 7) (2020-03-04) http://www.nhc.gov.cn/yzygj/s7653p/202003/46c9294a7dfe4cef80dc7f5912eb1989.shtml
[11] Corman VM, Landt O, Kaiser M, Molenkamp R, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020 pp. 1560-7917.
[12] LI Jin Ming. Real time Quantitative PCR Beijing Science Press, 2016.
Cite This Article
  • APA Style

    Shasha Zhang, Xiaoli Sun, Wei Guo, Jianfeng Xu. (2020). Simultaneous Detection of 2019 Novel Coronavirus and Influenza Virus by Double Fluorescent RT-PCR. American Journal of Laboratory Medicine, 5(1), 42-46. https://doi.org/10.11648/j.ajlm.20200501.16

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    ACS Style

    Shasha Zhang; Xiaoli Sun; Wei Guo; Jianfeng Xu. Simultaneous Detection of 2019 Novel Coronavirus and Influenza Virus by Double Fluorescent RT-PCR. Am. J. Lab. Med. 2020, 5(1), 42-46. doi: 10.11648/j.ajlm.20200501.16

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    AMA Style

    Shasha Zhang, Xiaoli Sun, Wei Guo, Jianfeng Xu. Simultaneous Detection of 2019 Novel Coronavirus and Influenza Virus by Double Fluorescent RT-PCR. Am J Lab Med. 2020;5(1):42-46. doi: 10.11648/j.ajlm.20200501.16

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  • @article{10.11648/j.ajlm.20200501.16,
      author = {Shasha Zhang and Xiaoli Sun and Wei Guo and Jianfeng Xu},
      title = {Simultaneous Detection of 2019 Novel Coronavirus and Influenza Virus by Double Fluorescent RT-PCR},
      journal = {American Journal of Laboratory Medicine},
      volume = {5},
      number = {1},
      pages = {42-46},
      doi = {10.11648/j.ajlm.20200501.16},
      url = {https://doi.org/10.11648/j.ajlm.20200501.16},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajlm.20200501.16},
      abstract = {This paper introduces a method for simultaneous detection of 2019 novel coronavirus (2019-nCoV) and influenza virus A and influenza virus B (FluA/B) by dual fluorescent RT-PCR, providing some references for the current clinical first-line practice against the epidemic. More than 12000 samples of nasopharyngeal swabs, sputum and anal swabs were collected. All samples were inactivated at 60°C for 40 min before testing, so as to protect medical examine. Nucleic acid detection kits from two manufacturers of 2019-nCoV and FluA/B were selected for the detection. Carboxyfluorescein (FAM) and green fluorescent protein (VIC) labeled probes were used to achieve simultaneous detection of the four gene targets using a double fluorescent RT-PCR reaction system. According to the results of nucleic acid detection, there is no cross infection between 2019-nCoV and FluA/B. The CT value of novel coronavirus nucleic acid in anal swab greater than CT value of sputum greater than CT value of nasopharyngeal swab in the same patient. This paper introduces a method for rapid and simultaneous detection of 2019-nCoV and FluA/B by dual fluorescent RT-PCR. It was improved the detection efficiency and reduce the cost, which could be used for rapid and emergent detection of 2019-nCoV and FluA/B. Our experiment is fast and accurate, could make reference for nucleic acid detection medical diagnosis for clinical medical staff.},
     year = {2020}
    }
    

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    AU  - Shasha Zhang
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    AB  - This paper introduces a method for simultaneous detection of 2019 novel coronavirus (2019-nCoV) and influenza virus A and influenza virus B (FluA/B) by dual fluorescent RT-PCR, providing some references for the current clinical first-line practice against the epidemic. More than 12000 samples of nasopharyngeal swabs, sputum and anal swabs were collected. All samples were inactivated at 60°C for 40 min before testing, so as to protect medical examine. Nucleic acid detection kits from two manufacturers of 2019-nCoV and FluA/B were selected for the detection. Carboxyfluorescein (FAM) and green fluorescent protein (VIC) labeled probes were used to achieve simultaneous detection of the four gene targets using a double fluorescent RT-PCR reaction system. According to the results of nucleic acid detection, there is no cross infection between 2019-nCoV and FluA/B. The CT value of novel coronavirus nucleic acid in anal swab greater than CT value of sputum greater than CT value of nasopharyngeal swab in the same patient. This paper introduces a method for rapid and simultaneous detection of 2019-nCoV and FluA/B by dual fluorescent RT-PCR. It was improved the detection efficiency and reduce the cost, which could be used for rapid and emergent detection of 2019-nCoV and FluA/B. Our experiment is fast and accurate, could make reference for nucleic acid detection medical diagnosis for clinical medical staff.
    VL  - 5
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Author Information
  • Panjin Centers for Disease Control/Panjin Center for Inspection and Testing, Panjin, China

  • Panjin Centers for Disease Control/Panjin Center for Inspection and Testing, Panjin, China

  • Panjin Centers for Disease Control/Panjin Center for Inspection and Testing, Panjin, China

  • Panjin Centers for Disease Control/Panjin Center for Inspection and Testing, Panjin, China

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