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

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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 102 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.

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Acknowledgments

This work was supported in part by the National Science and Technology Major Project (No. 2015ZX09102025), the Science and Technology Project of Zhejiang Province (2014C03001-2) and the Key Medical Subjects Construction Project of Zhejiang Province (XKQ-009-003). The authors thanks Dr. Gary Wong for English revision and helpful discussion of the manuscript.

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    Authors and Affiliations

    1. Key Laboratory of Emergency Detection for Public Health of Zhejiang Province, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China

      Changping Xu, Yan Feng & Yiyu Lu

    2. Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, China

      Hualei Wang, Hongli Jin, Na Feng, Xuexing Zheng, Zengguo Cao, Ling Li, Jianzhong Wang, Feihu Yan, Lina Wang, Hang Chi, Weiwei Gai, Chong Wang, Yongkun Zhao, Tiecheng Wang, Yuwei Gao, Songtao Yang & Xianzhu Xia

    3. Changchun SR Biological Technology Co., LTD, Changchun, 130012, China

      Hongli Jin

    4. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225000, China

      Hualei Wang, Na Feng, Xuexing Zheng, Yuwei Gao, Songtao Yang & Xianzhu Xia

    5. College of Animal Science, Henan Institute of Science and Technology, Xinxiang, 453003, China

      Jianzhong Wang

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    C. Xu and H. Wang contributed equally to this paper.

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    Xu, C., Wang, H., Jin, H. et al. Visual detection of Ebola virus using reverse transcription loop-mediated isothermal amplification combined with nucleic acid strip detection. Arch Virol 161, 1125–1133 (2016). https://doi.org/10.1007/s00705-016-2763-5

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