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| Pepin, Jean-francois; Riou, Antoine; Renault, Tristan. |
| Herpes and herpes-like virus infections have been reported in various marine mollusc species associated with high mortality rates. Following the characterisation and genome sequencing of ostreid herpesvirus 1 (OsHV-1), specific diagnostic tools have been developed based on conventional PCR techniques or in situ hybridisation. We have now developed a real-time PCR assay for rapid, sensitive and quantitative detection of OsHV-1, and compared it with a conventional PCR technique described previously. The new assay utilised SYBR® Green chemistry with specific primers C9/C10 targeting the C region. The melt curve analysis of OsHV-1 DNA or DNA extracted from infected material showed only one melting temperature peak (75.75 ± 0.1 °C). The assay had a detection... |
| Tipo: Text |
Palavras-chave: SYBR® Green; Real time PCR; Crassostrea gigas; Ostreid herpesvirus 1; Herpesvirus. |
| Ano: 2008 |
URL: http://archimer.ifremer.fr/doc/2008/publication-3951.pdf |
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| Riou, Antoine. |
| Depuis 1991, des épisodes de mortalités massives touchent les élevages ostréicoles français, notamment pour l'espèce Crassostrea gigas. Un des facteurs responsables de ces événements est le virus de l'Herpès de l'huître : Oyster Herpesvirus type 1 ou OsHV-1. Les travaux réalisés au cours de ce stage ont contribué dans un premier temps à l'amélioration du protocole d'analyse basé sur l'utilisation d'un appareil de Polymerase Chain Reaction (PCR) en temps résel, puis aux analyses et au suivi de la charge virale par cette technique, d'individus entrant dans le cadre des projets - TRIPLOMER et AQUAFIRST-UE. Cette approche permettra par la suite de réaliser des diganostics fiables de la maladie tout en précisant le nombre de copies ADN (Acide... |
| Tipo: Text |
Palavras-chave: Protocole; Herpes vrus; PCR quantitative; Huîtres creuses; Crassostrea gigas; Diagnostic. |
| Ano: 2006 |
URL: http://archimer.ifremer.fr/doc/00033/14459/11761.pdf |
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| Pepin, Jean-francois; Riou, Antoine; Renault, Tristan. |
| Herpes and herpes-like virus infections have been reported in various marine mollusc species associated with high mortality rates. Following the characterisation and genome sequencing of ostreid herpesvirus 1 (OsHV-1), specific diagnostic tools have been developed based on conventional PCR techniques or in situ hybridisation. We have now developed a real-time PCR assay for rapid, sensitive and quantitative detection of OsHV-1, and compared it with a conventional PCR technique described previously. The new assay utilised SYBR® Green chemistry with specific primers C9/C10 targeting the C region, which is present in two copies in the OsHV-1 genome and encodes two proteins of unknown function. The assay was applied successfully to rapid diagnosis (100 minutes)... |
| Tipo: Text |
Palavras-chave: Crassostrea gigas; PCR; Oyster; OsHV 1; Detection; Herpes virus 1. |
| Ano: 2007 |
URL: http://archimer.ifremer.fr/doc/2007/acte-3653.pdf |
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