A flow-FISH assay for the quantitative analysis of parvovirus B19 infected cells
作者： Elisabetta Manaresi , Gloria Bua , Francesca Bonvicini , Giorgio Gallinella
DOI: 10.1016/J.JVIROMET.2015.07.013
关键词: Flow cytometry 、 Biology 、 Flow-FISH 、 Molecular biology 、 Population 、 Fluorescence in situ hybridization 、 Parvovirus 、 Cell culture 、 Nucleic acid 、 Peripheral blood mononuclear cell
摘要: 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 differences in and investigate cell-virus interactions. Fluorescence situ hybridization (FISH) can be combined with flow cytometry (flow-FISH) enable detection of target nucleic acid sequences thousands individual cells short amount time. In present study, flow-FISH assay based use digoxigenin-labeled genomic probe has been developed discriminate B19V infected following vitro infection UT7/EpoS1 line EPCs (erythroid progenitor cells) generated from peripheral blood mononuclear cells. EPCs, viral acids were detected by starting 24 hpi up 48 hpi. The method, used together quantitative PCR techniques, very useful describe kinetics within heterogeneous population.
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