Quantitation of ranaviruses in cell culture and tissue samples

Riikka Holopainen, Jarno Honkanen, Britt Bang Jensen, Ellen Ariel, Hannele Tapiovaara

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    A quantitative real-time PCR (qPCR) based on a standard curve was developed for detection and quantitation of ranaviruses. The target gene for the qPCR was viral DNA polymerase (DNApol). All ten ranavirus isolates studied (Epizootic haematopoietic necrosis virus, EHNV; European catfish virus, ECV; European sheatfish virus, ESV; Frog virus 3, FV3; Bohle iridovirus, BIV; Doctor fish virus, DFV; Guppy virus 6, GV6; Pike-perch iridovirus, PPIV; Rana esculenta virus Italy 282/I02, REV282/I02 and Short-finned eel ranavirus, SERV) were detected with the qPCR assay. In addition, two fish cell lines – epithelioma papulosum cyprini (EPC) and bluegill fry (BF-2) – were infected with four of the isolates (EHNV, ECV, FV3 and DFV), and the viral quantity was determined from seven time points during the first three days after infection. The qPCR was also used to determine the viral load in tissue samples from pike (Esox lucius) fry challenged experimentally with EHNV.
    Original languageEnglish
    JournalJournal of Virological Methods
    Volume171
    Issue number1
    Pages (from-to)225-233
    ISSN0166-0934
    DOIs
    Publication statusPublished - 2011

    Keywords

    • Viral load
    • Ranavirus
    • DNA polymerase
    • Quantitative real-time PCR

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