A flow cytometric granularity assay for the quantification of infectious virus. Issue 47 (8th November 2019)

• Record Type: Journal Article
• Authors: Logan, Megan; Manalil, Julia; Notte, Christina; Kearse, Craig; George, Steve; Zeiser, Arno; Farrell, Patrick; Aucoin, Marc G.
• Is Part Of: Vaccine. Volume 37:Issue 47(2019)
• Page Start: 7090
• Page End: 7099
• Publication Date: 2019-11-08
• Subjects: Side-scatter -- Flow cytometry -- Herpes simplex virus -- Recombinant HSV-2 -- ALVAC -- Granularity; AV529-19 a Vero cell line designed specifically to express the UL5 and UL29 viral genes -- BHK-21 baby hamster kidney cells -- CCID50 cell culture infectious dose -- %CV coefficient of variation -- DI deionized water -- DMEM/F12 Dulbecco's Modified Eagle with Ham's F12 50:50 Mix Medium -- D-PBS Dulbecco's Phosphate Buffered Saline -- EDTA ethylenediaminetetraacetic acid -- FC flow cytometry-based assay -- HEK293 human embryonic kidney cells -- HI-FBS heat inactivated fetal bovine serum -- hpi hours post infection -- FSC forward scattered light -- LOD limit of detection -- MNA mouse neuroblastoma cells -- MOI multiplicity of infection -- PFU plaque forming units -- QT-35 quail muscle fibroblast cells -- rALVAC.gfp recombinant attenuated canarypox virus -- rHSV-2 recombinant herpes simplex virus serotype 2, also known as ACAM529 -- SSC side scattered light; Vaccines -- Periodicals; 615.372
• DOI: 10.1016/j.vaccine.2019.02.059
• Languages: English
• ISSNs: 0264-410X

Abstract

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 R 2 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.