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

Resultado de la investigación: Article

8 Citas (Scopus)

Resumen

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.

Idioma originalEnglish
Páginas (desde-hasta)291-294
Número de páginas4
PublicaciónJournal of Virological Methods
Volumen193
N.º2
DOI
EstadoPublished - 1 nov 2013

Huella dactilar

Hantavirus
Flow Cytometry
Virus Diseases
Viruses
Nucleocapsid Proteins
Virus Internalization
Endosomes
Viral Proteins
Infection
Antiviral Agents
Monoclonal Antibodies
Serum

ASJC Scopus subject areas

  • Virology

Citar esto

Barriga, G. P., Martínez-Valdebenito, C., Galeno, H., Ferrés, M., Lozach, P. Y., & Tischler, N. D. (2013). A rapid method for infectivity titration of Andes hantavirus using flow cytometry. Journal of Virological Methods, 193(2), 291-294. https://doi.org/10.1016/j.jviromet.2013.06.022
Barriga, Gonzalo P. ; Martínez-Valdebenito, Constanza ; Galeno, Héctor ; Ferrés, Marcela ; Lozach, Pierre Yves ; Tischler, Nicole D. / A rapid method for infectivity titration of Andes hantavirus using flow cytometry. En: Journal of Virological Methods. 2013 ; Vol. 193, N.º 2. pp. 291-294.
@article{4c782ec58c7e48718c4bea7f5e19833d,
title = "A rapid method for infectivity titration of Andes hantavirus using flow cytometry",
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, Hantavirus, Infectious virus titration, Nucleoprotein",
author = "Barriga, {Gonzalo P.} and Constanza Mart{\'i}nez-Valdebenito and H{\'e}ctor Galeno and Marcela Ferr{\'e}s and Lozach, {Pierre Yves} and Tischler, {Nicole D.}",
year = "2013",
month = "11",
day = "1",
doi = "10.1016/j.jviromet.2013.06.022",
language = "English",
volume = "193",
pages = "291--294",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier",
number = "2",

}

Barriga, GP, Martínez-Valdebenito, C, Galeno, H, Ferrés, M, Lozach, PY & Tischler, ND 2013, 'A rapid method for infectivity titration of Andes hantavirus using flow cytometry', Journal of Virological Methods, vol. 193, n.º 2, pp. 291-294. https://doi.org/10.1016/j.jviromet.2013.06.022

A rapid method for infectivity titration of Andes hantavirus using flow cytometry. / Barriga, Gonzalo P.; Martínez-Valdebenito, Constanza; Galeno, Héctor; Ferrés, Marcela; Lozach, Pierre Yves; Tischler, Nicole D.

En: Journal of Virological Methods, Vol. 193, N.º 2, 01.11.2013, p. 291-294.

Resultado de la investigación: Article

TY - JOUR

T1 - A rapid method for infectivity titration of Andes hantavirus using flow cytometry

AU - Barriga, Gonzalo P.

AU - Martínez-Valdebenito, Constanza

AU - Galeno, Héctor

AU - Ferrés, Marcela

AU - Lozach, Pierre Yves

AU - Tischler, Nicole D.

PY - 2013/11/1

Y1 - 2013/11/1

N2 - 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.

AB - 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.

KW - Flow cytometry

KW - Focus assay

KW - Hantavirus

KW - Infectious virus titration

KW - Nucleoprotein

UR - http://www.scopus.com/inward/record.url?scp=84880675422&partnerID=8YFLogxK

U2 - 10.1016/j.jviromet.2013.06.022

DO - 10.1016/j.jviromet.2013.06.022

M3 - Article

C2 - 23806566

AN - SCOPUS:84880675422

VL - 193

SP - 291

EP - 294

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

IS - 2

ER -

Barriga GP, Martínez-Valdebenito C, Galeno H, Ferrés M, Lozach PY, Tischler ND. A rapid method for infectivity titration of Andes hantavirus using flow cytometry. Journal of Virological Methods. 2013 nov 1;193(2):291-294. https://doi.org/10.1016/j.jviromet.2013.06.022