Rapid diagnosis and quantification of herpes simplex virus with a green fluorescent protein reporter system

A genetically modified cell line (Vero-ICP10-EGFP) was constructed for dete ction of herpes simplex virus (HSV) by a simple, rapid and direct method. T he cell line was developed by stable transfection of Vero cell with a plasm id encoding the green fluorescent protein (GFP) driven by the promoter of t he HSV-2 ICP10 gene. As early as 6 h after infection with HSV, fluorescence -emitting cells can be observed under a fluorescence microscope. A single i nfected cell emitting fluorescence can be observed with soft agar overlay b y inverted fluorescence microscopy. No induction of detectable fluorescence was seen following infections with human cytomegalovirus (HCMV), varicella tester virus (VZV), coxsackievirus A16 and enterovirus 71. Analysis by flo w cytometry also demonstrated that intensity of the triggered fluorescence is proportional to the titer of HSV inoculated. Taken together, this novel GFP reporter system could become a useful means for rapid detection and qua ntification of HSV in clinical specimens. (C) 2000 Elsevier Science B.V. Al l rights reserved.