Rapid identification and quantitation of cells infected by recombinant herpesvirus (pseudorabies virus) using a fluorescence-based beta-galactosidase assay and flow cytometry.
We recently described construction and use of a beta-galactosidase expression cassette in isolating recombinant pseudorabies virus (PrV) mutants (Mettenleiter and Rauh, 1990). We report here the identification and exact quantitation of cells infected by these mutants using an assay based on the reaction of intracellular beta-galactosidase expressed during infection by the recombinant viruses with the fluorogenic substrate fluorescein di-beta-D-galactopyranoside (FDG) followed by detection of positive cells in flow cytometry (FACS-Gal assay; Nolan et al., 1988). The detection method is fast, sensitive, and reliable, and yields quantitative results on single cell basis.
Saalmüller
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Federal Research Centre for Virus Diseases of Animals, Tübingen, Germany.
Mettenleiter
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TC
D000818:Animals; D001853:Bone Marrow / Q000382:microbiology*; D002417:Cattle; D002478:Cells, Cultured; D004848:Epithelium; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000382:microbiology*; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000032:analysis*; D013552:Swine; D001616:beta-Galactosidase / Q000032:analysis*
D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase