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Development of fast and sensitive protocols for the detection of viral pathogens using a small portable convection PCR platform

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Abstract

One of the most crucial steps for preventing viral pandemics is the early detection of the causative virus on site. Various molecular and immunological approaches have been developed for virus detection. In this study, we investigated the utility of the recently introduced convection polymerase chain reaction (cPCR) platform for the rapid and sensitive detection of various animal viruses in the field, including the foot-and-mouth disease virus (FMDV) and avian influenza viruses (AIVs). Primer sets were designed to simultaneously detect two highly conserved regions of the FMDV, including the 5′ untranslated region (5′-UTR) and 3D gene, and to specifically amplify the NP and hemagglutinin (HA) genes of H5 and H9 subtypes of AIVs. The portable cPCR system was able to amplify from as low as 1 to 10 copies of viral cDNAs in the singleplex mode and 10 to 100 copies of viral cDNAs in the duplex mode within 21 min. Thus, our data suggest that the cPCR protocols developed in this study are highly sensitive and enable quick detection of animal viruses in biological samples.

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Acknowledgements

This work was supported by the Research and Business Development Program of the Korea Institute for Advancement of Technology (Grant N0001697) and Korea Evaluation Institute of Industrial Technology (Grant 10080151) funded by the Ministry of Trade, Industry, and Energy, Korea.

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Authors and Affiliations

  1. Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, 790-784, Republic of Korea

    Myoung Hui Lee & Inhwan Hwang

  2. R&D Center, Ahram Biosystems Inc, Seoul, 133-120, Republic of Korea

    Kyung-Young Song & Hyun Jin Hwang

  3. Department of Biochemistry and Molecular Biology, College of Dentistry, Graduate School, Kyung Hee University, Seoul, 130-701, Republic of Korea

    Jeong Hee Kim

  4. Department of Life and Nanopharmaceutical Sciences, College of Dentistry, Graduate School, Kyung Hee University, Seoul, 130-701, Republic of Korea

    Jeong Hee Kim

  5. Department of Life Sciences, Pohang University of Science and Technology, Pohang, 790-784, Republic of Korea

    Inhwan Hwang

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  1. Myoung Hui Lee
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  2. Kyung-Young Song
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  3. Hyun Jin Hwang
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Correspondence to Jeong Hee Kim or Inhwan Hwang.

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Lee, M.H., Song, KY., Hwang, H.J. et al. Development of fast and sensitive protocols for the detection of viral pathogens using a small portable convection PCR platform. Mol Biol Rep 46, 5073–5077 (2019). https://doi.org/10.1007/s11033-019-04961-x

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  • DOI: https://doi.org/10.1007/s11033-019-04961-x

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