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

Gonzalo P. Barriga, Constanza Martínez-Valdebenito, Héctor Galeno, Marcela Ferrés, Pierre Yves Lozach, Nicole D. Tischler*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

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.

Original languageEnglish
Pages (from-to)291-294
Number of pages4
JournalJournal of Virological Methods
Volume193
Issue number2
DOIs
StatePublished - 2013
Externally publishedYes

Bibliographical note

Funding Information:
This work was financed by CONICYT through grant FONDECYT 1100756 and CONICYT basal funding PFB-16 .

ASJC Scopus subject areas

  • Virology

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