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Marine Biotechnology

, Volume 20, Issue 1, pp 35–44 | Cite as

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

  • Kuntong Jia
  • Yongming Yuan
  • Wei Liu
  • Lan Liu
  • Qiwei Qin
  • Meisheng Yi
Original Article

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.

Keywords

Singapore grouper iridovirus Antiviral drug screening Compound Cell viability-based assay Digital droplet PCR 

Notes

Funding Information

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).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

10126_2017_9785_MOESM1_ESM.docx (601 kb)
Table S1 (DOCX 600 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Zhuhai Key Laboratory of Marine Bioresources and Environment, School of Marine SciencesSun Yat-sen UniversityGuangzhouChina
  2. 2.Department of Biological SciencesNational University of SingaporeSingaporeSingapore
  3. 3.College of Marine SciencesSouth China Agricultural UniversityGuangzhouChina

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