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BMC Microbiol. 2014 Nov 14;14:271. doi: 10.1186/s12866-014-0271-x.

Development of reverse-transcription loop-mediated isothermal amplification assay for rapid detection of novel avian influenza A (H7N9) virus.

Author information

1
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. juanliu903@163.com.
2
Department of Transfusion Medicine, PLA Air Force General Hospital, Beijing, 100142, China. juanliu903@163.com.
3
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. nqgsucceed@163.com.
4
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. lj-pbs@163.com.
5
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. huyi_amms@163.com.
6
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. xiaofeng_li_bj@163.com.
7
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. susuzhai@163.com.
8
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. dengyq1977@126.com.
9
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. zhu_duck@163.com.
10
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. qingyu.zhu13@gmail.com.
11
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. qinede@sohu.com.
12
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. Jiang_tao@126.com.
13
Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No.20 Dongda Street, Beijing, Fengtai District, 100071, China. qincf@bmi.ac.cn.

Abstract

BACKGROUND:

The emerged human infection with avian influenza A (H7N9) virus in China since 2013 has aroused global concerns. There is great demand for simple and rapid diagnostic method for early detection of H7N9 to provide timely treatment and disease control. The aim of the current study was to develop a rapid, accurate and feasible reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of H7N9 virus.

RESULTS:

The detection limits of the H7- and N9-specific RT-LAMP assay were both approximately 0.2 PFU per reaction. No cross-reactivity was observed with other subtype of influenza viruses or common respiratory viral pathogens. The assay worked well with clinical specimens from patients and chickens, and exhibited high specificity and sensitivity.

CONCLUSIONS:

The H7/N9 specific RT-LAMP assay was sensitive and accurate, which could be a useful alternative in clinical diagnostics of influenza A (H7N9) virus, especially in the hospitals and laboratories without sophisticated diagnostic systems.

PMID:
25394781
PMCID:
PMC4234856
DOI:
10.1186/s12866-014-0271-x
[Indexed for MEDLINE]
Free PMC Article

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