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Rapid detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) by real-time, isothermal recombinase polymerase amplification assay

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Abstract

Infectious hypodermal and hematopoietic necrosis virus (IHHNV) causes mortality or runt deformity syndrome in penaeid shrimps and is responsible for significant economic losses in the shrimp aquaculture industry. Here, we describe a novel real-time isothermal recombinase polymerase amplification (RPA) assay developed for IHHNV detection. Using IHHNV plasmid standards and DNA samples from a variety of organisms, we evaluated the ability of the IHHNV-RPA assay to detect IHHNV based on analysis of its sensitivity, specificity, rapidity, and reproducibility. Probit analysis of eight independent experimental replicates indicated satisfactory performance of the RPA assay, which is sufficiently sensitive to detect as few as 4 copies of the IHHNV genome within 7 min at 39 °C with 95 % reliability. Therefore, this rapid RPA method has great potential for applications, either in field use or as a point of care diagnostic technique.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 41376135), the National High Technology Research and Development Program of China (863 Program, 2014AA093506), Doctoral Fund of Ministry of Education of China (20133104110006), Innovation Program of Shanghai Municipal Education Commission (14ZZ144), China, and Construction Program of Shanghai Committee of Science and Technology (11DZ2280300), China, and Marine Alliance for Science and Technology for Scotland.

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Authors and Affiliations

  1. Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation, Ministry of Agriculture, Shanghai, China

    Xiaoming Xia, Yongxin Yu, Linghao Hu, Yingjie Pan & Yongjie Wang

  2. Shanghai Engineering Research Center of Aquatic-Product Processing and Preservation, Shanghai, China

    Xiaoming Xia, Yongxin Yu, Yingjie Pan & Yongjie Wang

  3. College of Food Science and Technology, Shanghai Ocean University, Shanghai, China

    Xiaoming Xia, Yongxin Yu, Linghao Hu, Yingjie Pan, Shuling Yan & Yongjie Wang

  4. Institute of Aquaculture, University of Stirling, Stirling, UK

    Manfred Weidmann

  5. Institute of Biochemistry and Molecular Cell Biology, University of Göttingen, Göttingen, Germany

    Shuling Yan

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  1. Xiaoming Xia
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  2. Yongxin Yu
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Correspondence to Yongjie Wang.

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Xia, X., Yu, Y., Hu, L. et al. Rapid detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) by real-time, isothermal recombinase polymerase amplification assay. Arch Virol 160, 987–994 (2015). https://doi.org/10.1007/s00705-015-2357-7

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  • DOI: https://doi.org/10.1007/s00705-015-2357-7

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