A rapid method for infectivity titration of Andes hantavirus using flow cytometry.

Gonzalo P Barriga 1 Constanza Martínez-Valdebenito 2 Héctor Galeno 3 Marcela Ferrés 2 Pierre-Yves Lozach 4 Nicole D Tischler 1, 5, *
* Corresponding author
1 Molecular Virology Laboratory
2 Laboratorio de Infectología y Virología Molecular, División de Pediatría
3 Instituto de Salud Pública de Chile
4 INRS-IAF - Institut Armand Frappier
5 Facultad de Ciencias Biológicas
Abstract : The focus assay is currently the most commonly used technique for hantavirus titer determination. This method requires an incubation time of between 5 and 11 days to allow the appearance of foci after several rounds of viral infection. The following work presents a rapid Andes virus (ANDV) titration assay, based on viral nucleocapsid protein (N) detection in infected cells by flow cytometry. To this end, an anti-N monoclonal antibody was used that was developed and characterized previously. ANDV N could be detected as early as 6 h post-infection, while viral release was not observed until 24-48 h post-infection. Given that ANDV detection was performed during its first round of infection, a time reduction for titer determination was possible and provided results in only two days. The viral titer was calculated from the percentage of N positive cells and agreed with focus assay titers. Furthermore, the assay was applied to quantify the inhibition of ANDV cell entry by patient sera and by preventing endosome acidification. This novel hantavirus titration assay is a highly quantitative and sensitive tool that facilitates infectivity titration of virus stocks, rapid screening for antiviral drugs, and may be further used to detect and quantify infectious virus in human samples.
Keywords : Flow cytometry Focus assay Infectious virus titration Nucleoprotein Hantavirus
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Journal articles
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Submitted on : Wednesday, March 11, 2015 - 2:22:59 PM
Last modification on : Tuesday, August 13, 2019 - 2:30:11 PM

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Gonzalo P Barriga, Constanza Martínez-Valdebenito, Héctor Galeno, Marcela Ferrés, Pierre-Yves Lozach, et al.. A rapid method for infectivity titration of Andes hantavirus using flow cytometry.. Journal of Virological Methods, Elsevier, 2013, 193 (2), pp.291-4. ⟨10.1016/j.jviromet.2013.06.022⟩. ⟨pasteur-01130214⟩

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