Detection of a Novel Avian Influenza A (H7N9) Virus in Humans by Multiplex One-step Real-time RT-PCR Assay

Overview

Journal BMC Infect Dis

Publisher Biomed Central

Specialty Infectious Diseases

Date 2014 Oct 10

PMID 25298249

Citations 10

Authors

Jian Fan

David Cui

Siuying Lau

Guoliang Xie

Xichao Guo

Shufa Zheng

Xiaofeng Huang

Shigui Yang

Xianzhi Yang

Zhaoxia Huo

Fei Yu

Jianzhou Lou

Li Tian

Xuefen Li

Yuejiao Dong

Qiaoyun Zhu

Yu Chen

Affiliations

Soon will be listed here.

Abstract

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 (TCID50), 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.

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References

Ostrowsky B, Huang A, Terry W, Anton D, Brunagel B, Traynor L . Low pathogenic avian influenza A (H7N2) virus infection in immunocompromised adult, New York, USA, 2003. Emerg Infect Dis. 2012; 18(7):1128-31. PMC: 3376803. DOI: 10.3201/eid1807.111913. View

Chen Y, Liang W, Yang S, Wu N, Gao H, Sheng J . Human infections with the emerging avian influenza A H7N9 virus from wet market poultry: clinical analysis and characterisation of viral genome. Lancet. 2013; 381(9881):1916-25. PMC: 7134567. DOI: 10.1016/S0140-6736(13)60903-4. View

Zhang J, Feng Y, Hu D, Lv H, Zhu J, Cao M . Rapid and sensitive detection of H7N9 avian influenza virus by use of reverse transcription-loop-mediated isothermal amplification. J Clin Microbiol. 2013; 51(11):3760-4. PMC: 3889764. DOI: 10.1128/JCM.01907-13. View

Ge Y, Wu B, Qi X, Zhao K, Guo X, Zhu Y . Rapid and sensitive detection of novel avian-origin influenza A (H7N9) virus by reverse transcription loop-mediated isothermal amplification combined with a lateral-flow device. PLoS One. 2013; 8(8):e69941. PMC: 3731295. DOI: 10.1371/journal.pone.0069941. View

Gao H, Lu H, Cao B, Du B, Shang H, Gan J . Clinical findings in 111 cases of influenza A (H7N9) virus infection. N Engl J Med. 2013; 368(24):2277-85. DOI: 10.1056/NEJMoa1305584. View

Corman V, Eickmann M, Landt O, Bleicker T, Brunink S, Eschbach-Bludau M . Specific detection by real-time reverse-transcription PCR assays of a novel avian influenza A(H7N9) strain associated with human spillover infections in China. Euro Surveill. 2013; 18(16):20461. View

Cheng V, Chan J, Wen X, Wu W, Que T, Chen H . Infection of immunocompromised patients by avian H9N2 influenza A virus. J Infect. 2011; 62(5):394-9. DOI: 10.1016/j.jinf.2011.02.007. View

Li Q, Zhou L, Zhou M, Chen Z, Li F, Wu H . Epidemiology of human infections with avian influenza A(H7N9) virus in China. N Engl J Med. 2013; 370(6):520-32. PMC: 6652192. DOI: 10.1056/NEJMoa1304617. View

Zhang Y, Mao H, Yan J, Wang X, Zhang L, Guus K . Development of novel AllGlo-probe-based one-step multiplex qRT-PCR assay for rapid identification of avian influenza virus H7N9. Arch Virol. 2014; 159(7):1707-13. DOI: 10.1007/s00705-014-1979-5. View

10.

Kudo K, Binh N, Manabe T, Co D, Tuan N, Izumi S . Clinical preparedness for severe pneumonia with highly pathogenic avian influenza A (H5N1): experiences with cases in Vietnam. Respir Investig. 2012; 50(4):140-50. DOI: 10.1016/j.resinv.2012.08.005. View

11.

Dong H, Zhang Y, Xiong H, Yan A, Ding G, Chen Y . Detection of human novel influenza A (H1N1) viruses using multi-fluorescent real-time RT-PCR. Virus Res. 2009; 147(1):85-90. DOI: 10.1016/j.virusres.2009.10.011. View

12.

Wong C, Zhu H, Li O, Leung Y, Chan M, Guan Y . Molecular detection of human H7N9 influenza A virus causing outbreaks in China. Clin Chem. 2013; 59(7):1062-7. DOI: 10.1373/clinchem.2013.208975. View

13.

Zhu Z, Fan H, Qi X, Qi Y, Shi Z, Wang H . Development and evaluation of a SYBR green-based real time RT-PCR assay for detection of the emerging avian influenza A (H7N9) virus. PLoS One. 2013; 8(11):e80028. PMC: 3835827. DOI: 10.1371/journal.pone.0080028. View

14.

Chen H, Yuan H, Gao R, Zhang J, Wang D, Xiong Y . Clinical and epidemiological characteristics of a fatal case of avian influenza A H10N8 virus infection: a descriptive study. Lancet. 2014; 383(9918):714-21. DOI: 10.1016/S0140-6736(14)60111-2. View

15.

Lu S, Xi X, Zheng Y, Cao Y, Liu X, Lu H . Analysis of the clinical characteristics and treatment of two patients with avian influenza virus (H7N9). Biosci Trends. 2013; 7(2):109-12. View

16.

Liu T, Bi Z, Wang X, Li Z, Ding S, Bi Z . One family cluster of avian influenza A(H7N9) virus infection in Shandong, China. BMC Infect Dis. 2014; 14:98. PMC: 3974040. DOI: 10.1186/1471-2334-14-98. View

17.

Gao R, Cao B, Hu Y, Feng Z, Wang D, Hu W . Human infection with a novel avian-origin influenza A (H7N9) virus. N Engl J Med. 2013; 368(20):1888-97. DOI: 10.1056/NEJMoa1304459. View

18.

Guan Y, Farooqui A, Zhu H, Dong W, Wang J, Kelvin D . H7N9 Incident, immune status, the elderly and a warning of an influenza pandemic. J Infect Dev Ctries. 2013; 7(4):302-7. DOI: 10.3855/jidc.3675. View

19.

Nie K, Zhao X, Ding X, Li X, Zou S, Guo J . Visual detection of human infection with influenza A (H7N9) virus by subtype-specific reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye. Clin Microbiol Infect. 2013; 19(8):E372-5. DOI: 10.1111/1469-0691.12263. View

20.

Kim H, Park C, Lee Y, Oem J, Kang H, Choi J . Low pathogenic H7 subtype avian influenza viruses isolated from domestic ducks in South Korea and the close association with isolates of wild birds. J Gen Virol. 2012; 93(Pt 6):1278-1287. DOI: 10.1099/vir.0.041269-0. View