Fluorescence-based antiviral assay for the evaluation of compounds against vaccinia virus, varicella zoster virus and human cytomegalovirus

被引：22

作者：

Dal Pozzo, Fabiana ^{[1
,2
]}

Andrei, Graciela ^{[1
]}

Daelemans, Dirk ^{[1
]}

Winkler, Michael ^{[3
]}

Piette, Jacques ^{[4
]}

De Clercq, Erik ^{[1
]}

Snoeck, Robert ^{[1
]}

机构：

[1] Katholieke Univ Leuven, Rega Inst Med Res, B-3000 Louvain, Belgium

[2] Alma Mater Studiorum, Dept Vet Publ Hlth & Anim Pathol, I-40064 Ozzano Dell Emilia, Bologna, Italy

[3] Univ Ulm, Inst Virol, D-89081 Ulm, Germany

[4] GIGA Res, Virol & Immunol Unit, B-4000 Liege, Belgium

来源：

JOURNAL OF VIROLOGICAL METHODS | 2008年 / 151卷 / 01期

关键词：

vaccinia virus; varicella zoster virus; human cytomegalovirus; GFP-recombinant virus; antiviral assay; fluorescence-based antiviral assay;

D O I：

10.1016/j.jviromet.2008.03.025

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Recombinant vaccinia virus (VACV), varicella zoster virus (VZV) and two human cytomegaloviruses (HCMV) expressing the green fluorescent protein (GFP) and the enhanced yellow fluorescent protein (EYFP) were used to develop a fluorescence-based assay for testing antiviral compounds. Infection of human embryonic lung fibroblasts (HEL) with the different recombinant viruses produced stable and detectable amount of GFP and EYFP signal as quantitated by automated fluorometry. The sensitivity of the recombinant viruses to a panel of antiviral drugs was measured and the fluorescence-based assay was compared to the cytopathic effect reduction assay (CPE-RA) in case of VACV and HCMV or to the plaque reduction assay (PRA) in case of VZV. The 50% inhibitory concentration (IC50) values for reference anti-pox and anti-herpesvirus compounds were comparable to those determined by CPE-RA or PRA assays. Furthermore the fluorimetric data could be confirmed by a flow cytometry assay. GFP- and EYFP-recombinant viruses proved to be a convenient tool for the evaluation of antiviral agents. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：66 / 73

页数：8

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