Design and validation of a RT-qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers

© 2016 John Wiley & Sons Ltd.

Bibliographische Detailangaben
Veröffentlicht in:Journal of fish diseases. - 1998. - 40(2017), 9 vom: 01. Sept., Seite 1155-1167
1. Verfasser: Vázquez, D (VerfasserIn)
Weitere Verfasser: Cutrín, J M, Olveira, J G, Dopazo, C P
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2017
Zugriff auf das übergeordnete Werk:Journal of fish diseases
Schlagworte:Journal Article IPNV RT-qPCR diagnosis quantification DNA Primers
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245 1 0 |a Design and validation of a RT-qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers 
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520 |a Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT-qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT-qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID50  mL-1 , 50 pfu mL-1 or 66 RNA copies mL-1 , depending on the standard. All the standard curves showed high reliability (R2  > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV 
650 4 |a Journal Article 
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700 1 |a Cutrín, J M  |e verfasserin  |4 aut 
700 1 |a Olveira, J G  |e verfasserin  |4 aut 
700 1 |a Dopazo, C P  |e verfasserin  |4 aut 
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