18 research outputs found

    Nonspecific Cellular and Humoral Defence Mechanisms in Sheatfish (Silurus glanis)

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    Anti-Birnavirus Activity of Methisoprinol – in vitro Study with Infectious Pancreatic Necrosis Virus (IPNV)

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    This study was conducted to evaluate the influence of methisoprinol, synthetic anti-viral product, on the IPNV replication in vitro by measuring viral RNA synthesis. The monolayers of RTG-2 cells in tissue culture plates (Multiwell, 24 wells, Becton Dickinson, USA) wa

    Detection of Echinococcus multilocularis in wild boars in France using PCR techniques against larval form

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    Recently, new data have been collected on the distribution and ecology of Echinococcus multilocularis in European countries. Different ungulates species such as pig, goat, sheep, cattle and horse are known to host incomplete development of larval E. multilocularis. We report a case of E. multilocularis portage in two wild boars from a high endemic area in France (Department of Jura). Histological examination was performed and the DNA was isolated from hepatic lesions then amplified by using three PCR methods in two distinct institutes. Molecular characterisation of PCR products revealed 99% nucleotide sequence homology with the specific sequence of the U1 sn RNA gene of E. multilocularis, 99 and 99.9% nucleotide sequence homology with the specific sequence of the cytochrome oxydase gene of Echinococcus genus and 99.9% nucleotide sequence homology with a genomic DNA sequence of Echinococcus genus for the first and the second wild boar, respectively. (c) 2004 Elsevier B.V. All rights reserved

    Differentiation between Cyprinid herpesvirus type-3 lineages using duplex PCR

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    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

    Development of real-time PCR assays for the detection and differentiation of Australian and European ranaviruses.

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    Serious systemic disease in fish and amphibians is associated with the ranaviruses, epizootic haernatopoietic necrosis virus (EHNV) and Bohle iridovirus (BIV) in Australia, and European sheattish virus (ESV) and European catfish virus (ECV) in Europe. EHNV, ESV and ECV are recognized causative agents of the OlE (Office International des Epizooties) notifiable systemic necrotizing iridovirus syndrome and are currently identified by proteinbased assays, none of which are able to rapidly identify the specific agents. The aim of this study was to develop T aqMan real-time PCR assays that differentiated these viruses using nucleotide sequence variation in two ranavirus genes. A conserved probe representing 100% sequence homology was used as a reference for virus-specific probes. The virus-specific probes produced a similar signal level to the conserved probe while those probes binding to non-target viral DNA produced an altered fluorescent curve. The pattern of probe binding was characteristic for each virus. Sensitivity, specificity and dynamic range of the assay were assessed. The test is currently useful as a research and initial screening tool, with the potential to become a sensitive and specific method for detection and differentiation of ranaviruses with further development
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