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Arch Virol. 2016 May;161(5):1125-33. doi: 10.1007/s00705-016-2763-5. Epub 2016 Jan 30.

Visual detection of Ebola virus using reverse transcription loop-mediated isothermal amplification combined with nucleic acid strip detection.

Xu C1, Wang H2,3, Jin H2,4, Feng N2,3, Zheng X2,3, Cao Z2, Li L2, Wang J2,5, Yan F2, Wang L2, Chi H2, Gai W2, Wang C2, Zhao Y2, Feng Y1, Wang T2, Gao Y2,3, Lu Y6, Yang S7,8, Xia X9,10.

Author information

1
Key Laboratory of Emergency Detection for Public Health of Zhejiang Province, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China.
2
Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, China.
3
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225000, China.
4
Changchun SR Biological Technology Co., LTD, Changchun, 130012, China.
5
College of Animal Science, Henan Institute of Science and Technology, Xinxiang, 453003, China.
6
Key Laboratory of Emergency Detection for Public Health of Zhejiang Province, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China. luyiyuzjh@163.com.
7
Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, China. yst62041@163.com.
8
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225000, China. yst62041@163.com.
9
Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, China. xiaxzh@cae.cn.
10
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225000, China. xiaxzh@cae.cn.

Abstract

Ebola virus (species Zaire ebolavirus) (EBOV) is highly virulent in humans. The largest recorded outbreak of Ebola hemorrhagic fever in West Africa to date was caused by EBOV. Therefore, it is necessary to develop a detection method for this virus that can be easily distributed and implemented. In the current study, we developed a visual assay that can detect EBOV-associated nucleic acids. This assay combines reverse transcription loop-mediated isothermal amplification and nucleic acid strip detection (RT-LAMP-NAD). Nucleic acid amplification can be achieved in a one-step process at a constant temperature (58 °C, 35 min), and the amplified products can be visualized within 2-5 min using a nucleic acid strip detection device. The assay is capable of detecting 30 copies of artificial EBOV glycoprotein (GP) RNA and RNA encoding EBOV GP from 10(2) TCID50 recombinant viral particles per ml with high specificity. Overall, the RT-LAMP-NAD method is simple and has high sensitivity and specificity; therefore, it is especially suitable for the rapid detection of EBOV in African regions.

PMID:
26831931
DOI:
10.1007/s00705-016-2763-5
[Indexed for MEDLINE]

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