FN Archimer Export Format
PT J
TI Rapid and sensitive detection of ostreid herpesvirus 1 in oyster samples by real-time PCR
BT 
AF PEPIN, Jean-Francois
   RIOU, Antoine
   RENAULT, Tristan
AS 1:1;2:1;3:1;
FF 1:PDG-DOP-DCN-AGSAE-LGP;2:PDG-DOP-DCN-AGSAE-LGP;3:PDG-DOP-DCN-AGSAE-LGP;
C1 IFREMER, LGP, F-17390 La Tremblade, France.
C2 IFREMER, FRANCE
SI LA TREMBLADE
SE PDG-DOP-DCN-AGSAE-LGP
IN WOS Ifremer jusqu'en 2018
IF 2.077
TC 127
UR https://archimer.ifremer.fr/doc/2008/publication-3951.pdf
LA English
DT Article
DE ;SYBR® Green;Real time PCR;Crassostrea gigas;Ostreid herpesvirus 1;Herpesvirus
AB 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 limit of 4 copies/μL of viral genomic DNA and a dynamic range of 5 logs. Using infected oyster samples as template, the assay was about 100-fold more sensitive than single PCR method using C2/C6 primers. The assay was applied successfully for rapid diagnosis (100 min) and quantitation of OsHV-1 in different developmental stages of Crassostrea gigas. Although it already exists a competitive PCR method to quantify OsHV-1 DNA, quantitative data that will emerge in future using the new sensitive and reliable assay will illuminate aspects of pathogenesis, in particular the viral loads in asymptomatic oysters and the kinetics of infection in specific target tissues.
PY 2008
PD MAY
SO Journal of Virological Methods
SN 0166-0934
PU Elsevier
VL 149
IS 2
UT 000255622400011
BP 269
EP 276
DI 10.1016/j.jviromet.2008.01.022
ID 3951
ER

EF