title: Detection of Tilapia Lake Virus in Clinical Samples by Culturing and Nested Reverse Transcription-PCR.
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.
authors.affiliation: Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Japhette Esther
authors.initials: JE
authors.lastname: Kembou Tsofack
authors.affiliation: Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Rachel
authors.initials: R
authors.lastname: Zamostiano
authors.affiliation: Fish Diseases Laboratory, The Kimron Veterinary Institute, Bet Dagan, Israel.
authors.forename: Salsabeel
authors.initials: S
authors.lastname: Watted
authors.affiliation: Fish Diseases Laboratory, The Kimron Veterinary Institute, Bet Dagan, Israel.
authors.forename: Asaf
authors.initials: A
authors.lastname: Berkowitz
authors.affiliation: Fish Diseases Laboratory, The Kimron Veterinary Institute, Bet Dagan, Israel.
authors.forename: Ezra
authors.initials: E
authors.lastname: Rosenbluth
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: Nischay
authors.initials: N
authors.lastname: Mishra
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Briese
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: W Ian
authors.initials: WI
authors.lastname: Lipkin
authors.affiliation: Marine Medicine Program, Pathobiology, School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Kabuusu
authors.affiliation: Marine Medicine Program, Pathobiology, School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Hugh
authors.initials: H
authors.lastname: Ferguson
authors.affiliation: Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, Scotland.
authors.forename: Jorge
authors.initials: J
authors.lastname: Del Pozo
authors.affiliation: Fish Diseases Laboratory, The Kimron Veterinary Institute, Bet Dagan, Israel eldar@int.gov.il eranbac@post.tau.ac.il.
authors.forename: Avi
authors.initials: A
authors.lastname: Eldar
authors.affiliation: Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel eldar@int.gov.il eranbac@post.tau.ac.il.
authors.forename: Eran
authors.initials: E
authors.lastname: Bacharach
mesh_terms: D000818:Animals; D002460:Cell Line; D003105:Colombia; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology; D009975:Orthomyxoviridae / Q000302:isolation & purification*; D009976:Orthomyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017210:Tilapia / Q000821:virology*; D014776:Virus Cultivation / Q000379:methods*
chemical_list: D012367:RNA, Viral
keywords: PCR; TiLV; diagnosis; tilapia; virus