A flow-FISH assay for the quantitative analysis of parvovirus B19 infected cells.

Abstract

Human parvovirus B19 (B19V) replication is a process highly dependent on the cellular environment, therefore methodologies allowing for analysis at single cell level could represent effective tools to understand cell-to cell differences in the replication process and to investigate cell-virus interactions. Fluorescence in situ hybridization (FISH) can be combined with flow cytometry (flow-FISH) to enable the detection of target nucleic acid sequences in thousands of individual cells in a short amount of time. In the present study, a flow-FISH assay based on the use of a digoxigenin-labeled genomic probe has been developed to discriminate B19V infected cells following in vitro infection of UT7/EpoS1 cell line and EPCs (erythroid progenitor cells) generated from peripheral blood mononuclear cells. In B19V infected UT7/EpoS1 and EPCs, viral nucleic acids were detected by the flow-FISH assay starting from 24 hpi up to 48 hpi. The method, used together with quantitative PCR techniques, can be very useful to describe the kinetics of B19V infection within a heterogeneous cell population.

• Authors: Elisabetta Manaresi, Gloria Bua, Francesca Bonvicini, Giorgio Gallinella
• Journal: Journal of virological methods (J Virol Methods) Vol. 223 Pg. 50-4 (Oct 2015) ISSN: 1879-0984 [Electronic] Netherlands
• PMID: 26231787 (Publication Type: Journal Article)
• Copyright: Copyright © 2015 Elsevier B.V. All rights reserved.

Topics

• Cells, Cultured
• Flow Cytometry (methods)
• Humans
• In Situ Hybridization, Fluorescence (methods)
• Parvovirus B19, Human (growth & development)
• Virology (methods)