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Article

Quantitative Flow Cytometry to Measure Viral Production Using Infectious Pancreatic Necrosis Virus as a Model: A Preliminary Study

Instituto de Acuicultura, Departamento de Microbiología, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain
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Appl. Sci. 2018, 8(10), 1734; https://doi.org/10.3390/app8101734
Received: 6 September 2018 / Revised: 22 September 2018 / Accepted: 22 September 2018 / Published: 26 September 2018
(This article belongs to the Section Applied Biosciences and Bioengineering)
In recent decades, flow cytometry (FCM) has become an important tool in virology, due to its applications in viral replication and viral-cell interactions, as well as its capacity to quantify proteins (qFCM). In the present study, we have designed and evaluated a qFCM procedure for the in vitro analysis and quantification of fish viral proteins, using the infectious pancreatic necrosis virus (IPNV) as a model. We have also tested its use for viral titration and adapted the MARIS (method for analysing RNA following intracellular sorting) method for simultaneous quantification of viral RNA expression in infected cells. The procedure has proved to be repeatable and reproducible to an acceptable level, although to ensure reproducibility, the repetition of standard curves is inevitable. Regarding its use for viral quantification, a direct relationship (by a second-degree polynomial regression) between viral titres and Molecules of Equivalent Soluble Fluorochrome (MESF) was observed. Finally, the results support the use of this technology, not only for virus quantification, but also to study viral replication from a quantitative approach. View Full-Text
Keywords: qFCM; IPNV; viral replication qFCM; IPNV; viral replication
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MDPI and ACS Style

Vázquez, D.; López-Vázquez, C.; Olveira, J.G.; Bandín, I.; Dopazo, C.P. Quantitative Flow Cytometry to Measure Viral Production Using Infectious Pancreatic Necrosis Virus as a Model: A Preliminary Study. Appl. Sci. 2018, 8, 1734. https://doi.org/10.3390/app8101734

AMA Style

Vázquez D, López-Vázquez C, Olveira JG, Bandín I, Dopazo CP. Quantitative Flow Cytometry to Measure Viral Production Using Infectious Pancreatic Necrosis Virus as a Model: A Preliminary Study. Applied Sciences. 2018; 8(10):1734. https://doi.org/10.3390/app8101734

Chicago/Turabian Style

Vázquez, Diego, Carmen López-Vázquez, José G. Olveira, Isabel Bandín, and Carlos P. Dopazo 2018. "Quantitative Flow Cytometry to Measure Viral Production Using Infectious Pancreatic Necrosis Virus as a Model: A Preliminary Study" Applied Sciences 8, no. 10: 1734. https://doi.org/10.3390/app8101734

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