A flow cytometric granularity assay for the quantification of infectious virus

Megan Logan; Julia Manalil; Christina Notte; Craig Kearse; Steve George; Arno Zeiser; Patrick Farrell; Marc G Aucoin

doi:10.1016/j.vaccine.2019.02.059

A flow cytometric granularity assay for the quantification of infectious virus

Vaccine. 2019 Nov 8;37(47):7090-7099. doi: 10.1016/j.vaccine.2019.02.059. Epub 2019 Apr 16.

Authors

Megan Logan¹, Julia Manalil¹, Christina Notte², Craig Kearse², Steve George², Arno Zeiser², Patrick Farrell², Marc G Aucoin³

Affiliations

¹ Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L3G1, Canada.
² Sanofi Pasteur, 1755 Steeles Ave. West, Toronto, Ontario M2R 3T4, Canada.
³ Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L3G1, Canada. Electronic address: maucoin@uwaterloo.ca.

PMID: 31630940
DOI: 10.1016/j.vaccine.2019.02.059

Abstract

A flow cytometry-based assay was developed to assess the infective titer of two recombinant viruses: a recombinant herpes simplex type 2 (rHSV-2) and a recombinant canary pox (rALVAC.gfp). This method uses granularity of infected Vero and QT-35 cells, respectively, and correlates this to the infectious titer of virus samples. The percent of the cell populations with a high level of granularity could accurately be correlated to viral titers obtained through a traditional plaque assay, with R² values greater than 0.8 using a semi-logarithmic scale. This approach offers a rapid, high-throughput method for infectious virus titration with similar accuracy to a traditional plaque assay.

Keywords: ALVAC; Flow cytometry; Granularity; Herpes simplex virus; Recombinant HSV-2; Side-scatter.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Chlorocebus aethiops
Flow Cytometry / methods*
Herpesvirus 2, Human / isolation & purification*
Vero Cells
Viral Load / methods
Viral Plaque Assay / methods
Virus Diseases / virology*