Saalmüller A, Mettenleiter T C
Federal Research Centre for Virus Diseases of Animals, Tübingen, Germany.
J Virol Methods. 1993 Sep;44(1):99-108. doi: 10.1016/0166-0934(93)90012-g.
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.
我们最近描述了一种β-半乳糖苷酶表达盒的构建及其在分离重组伪狂犬病病毒（PrV）突变体中的应用（Mettenleiter和Rauh，1990）。在此，我们报告使用一种基于重组病毒感染期间表达的细胞内β-半乳糖苷酶与荧光底物二-β-D-吡喃半乳糖苷荧光素（FDG）反应的检测方法来鉴定和精确定量感染这些突变体的细胞，随后通过流式细胞术检测阳性细胞（FACS-Gal检测法；Nolan等人，1988）。该检测方法快速、灵敏且可靠，可在单细胞基础上产生定量结果。
