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Identification of Inhibitory Compounds Against Singapore Grouper Iridovirus Infection by Cell Viability-Based Screening Assay and Droplet Digital PCR

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Abstract

Singapore grouper iridovirus (SGIV) is one of the major causative agents of fish diseases and has caused significant economic losses in the aquaculture industry. There is currently no commercial vaccine or effective antiviral treatment against SGIV infection. Annually, an increasing number of small molecule compounds from various sources have been produced, and many are proved to be potential inhibitors against viruses. Here, a high-throughput in vitro cell viability-based screening assay was developed to identify antiviral compounds against SGIV using the luminescent-based CellTiter-Glo reagent in cultured grouper spleen cells by quantificational measurement of the cytopathic effects induced by SGIV infection. This assay was utilized to screen for potential SGIV inhibitors from five customized compounds which had been reported to be capable of inhibiting other viruses and 30 compounds isolated from various marine organisms, and three of them [ribavirin, harringtonine, and 2-hydroxytetradecanoic acid (2-HOM)] were identified to be effective on inhibiting SGIV infection, which was further confirmed with droplet digital PCR (ddPCR). In addition, the ddPCR results revealed that ribavirin and 2-HOM inhibited SGIV replication and entry in a dose-dependent manner, and harringtonine could reduce SGIV replication rather than entry at the working concentration without significant toxicity. These findings provided an easy and reliable cell viability-based screening assay to identify compounds with anti-SGIV effect and a way of studying the anti-SGIV mechanism of compounds.

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Funding

This work was supported by the National Natural Science Foundation of China (31502195, 31602191), the Zhuhai Scholar Professor Program (2015), the Guangdong Natural Science Foundation (2015A030308012), and the Science and Technology Planning Project of Guangdong Province (2017A030303010).

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Author notes

  1. Kuntong Jia and Yongming Yuan contributed equally to this work.

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Zhuhai Key Laboratory of Marine Bioresources and Environment, School of Marine Sciences, Sun Yat-sen University, Guangzhou, Guangdong, China

    Kuntong Jia, Wei Liu, Lan Liu & Meisheng Yi

  2. Department of Biological Sciences, National University of Singapore, Science Drive 4, Singapore, 117543, Singapore

    Yongming Yuan

  3. College of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China

    Qiwei Qin

Authors
  1. Kuntong Jia
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  2. Yongming Yuan
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  3. Wei Liu
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  4. Lan Liu
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  5. Qiwei Qin
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  6. Meisheng Yi
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Correspondence to Qiwei Qin or Meisheng Yi.

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Jia, K., Yuan, Y., Liu, W. et al. Identification of Inhibitory Compounds Against Singapore Grouper Iridovirus Infection by Cell Viability-Based Screening Assay and Droplet Digital PCR. Mar Biotechnol 20, 35–44 (2018). https://doi.org/10.1007/s10126-017-9785-1

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