Differentiation between Cyprinid herpesvirus type-3 lineages using duplex PCR

L. Bigarre, M. Baud, J. Cabon, J. Antychowicz, S.M. Bergmann, M.Y. Engelsma, F. Pozet, M. Reichert, J. Castric

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    Abstract

    To date, all the isolates of Cyprinid herpesvirus type-3 (CyHV3) responsible for serious outbreaks in carps Cyprinus carpio have been found to be very similar or identical on the basis of DNA sequences of a few reference genes. However, two genetic lineages (U/I and J) are distinguished by full-length genome sequencing. Two molecular markers presenting genetic variations were targeted for developing a duplex PCR assay able to distinguish CyHV3-U/I from CyHV3-J while avoiding DNA sequencing. The method was validated on a series of 42 samples of infected carps from France, The Netherlands and Poland collected from 2001 to 2008. Among these samples, both the U/I and J genotypes were identified, but also a third genotype representing a genetic intermediate between U/I and J for one of the two molecular markers. A classification of CyHV3 genotypes, based on the alleles of the two molecular markers, is proposed. The assay is easy to perform and provides a genotype information with samples moderately or highly concentrated. This tool should improve our knowledge regarding the present distribution and future diversification of this emerging virus.
    Original languageEnglish
    Pages (from-to)51-57
    JournalJournal of Virological Methods
    Volume158
    Issue number1-2
    DOIs
    Publication statusPublished - 2009

    Keywords

    • simplex-virus type-1
    • polymerase-chain-reaction
    • koi herpesvirus
    • common carp
    • thymidine kinase
    • infected fish
    • dna
    • mortality
    • disease
    • tissues

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