Development of reverse-transcription loop-mediated isothermal amplification assay for rapid detection of novel avian influenza A (H7N9) virus.
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.
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.|Department of Transfusion Medicine, PLA Air Force General Hospital, Beijing, 100142, China. juanliu903@163.com.
Juan
J
Liu
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.
Qing-Gong
QG
Nian
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.
Jing
J
Li
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.
Yi
Y
Hu
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.
Xiao-Feng
XF
Li
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.
Yu
Y
Zhang
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.
Yong-Qiang
YQ
Deng
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.
Shun-Ya
SY
Zhu
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.
Qing-Yu
QY
Zhu
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.
E-De
ED
Qin
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.
Tao
T
Jiang
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.
Cheng-Feng
CF
Qin
D000818:Animals; D002645:Chickens; D002681:China; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods