Detection of tilapia lake virus in clinical samples by culturing and nested reverse transcription-PCR

Japhette Esther Kembou Tsofack, Rachel Zamostiano, Salsabeel Watted, Asaf Berkowitz, Ezra Rosenbluth, Nischay Mishra, Thomas Briese, W. Ian Lipkin, Richard M. Kabuusu, Hugh Ferguson, Jorge Del Pozo, Avi Eldar, Eran Bacharach*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Tilapia are an important group of farmed fish that serve as a significant protein source worldwide. In recent years, substantial mortality of wild tilapia has been observed in the Sea of Galilee and in commercial ponds in Israel and Ecuador. We have identified the etiological agent of these mass die-offs as a novel orthomyxo-like virus and named it tilapia lake virus (TiLV). Here, we provide the conditions for efficient isolation, culturing, and quantification of the virus, including the use of susceptible fish cell lines. Moreover, we describe a sensitive nested reverse transcription- PCR (RT-PCR) assay allowing the rapid detection of TiLV in fish organs. This assay revealed, for the first time to our knowledge, the presence of TiLV in diseased Colombian tilapia, indicating a wider distribution of this emerging pathogen and stressing the risk that TiLV poses for the global tilapia industry. Overall, the described procedures should provide the tilapia aquaculture industry with important tools for the detection and containment of this pathogen.

Original languageEnglish
Pages (from-to)759-767
Number of pages9
JournalJournal of Clinical Microbiology
Volume55
Issue number3
DOIs
StatePublished - Mar 2017

Funding

FundersFunder number
Israel Ministry of Agriculture & Rural Development Chief Scientist Office847-0389-14
United States - Israel Binational Agricultural Research and Development FundIS-4903-16C, BARD IS-4583-13
Tel Aviv University

    Keywords

    • Diagnosis
    • PCR
    • Tilapia
    • Tilv
    • Virus

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