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 detection of herpes simplex virus (HSV) by a simple, rapid and direct method. The cell line was developed by stable transfection of Vero cell with a plasmid encoding the green fluorescent protein (GFP) driven by the promoter of the 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 infected cell emitting fluorescence can be observed with soft agar overlay by inverted fluorescence microscopy. No induction of detectable fluorescence was seen following infections with human cytomegalovirus (HCMV), varicella zoster virus (VZV), coxsackievirus A16 and enterovirus 71. Analysis by flow 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 quantification of HSV in clinical specimens.
Kung
S H
SH
Faculty of Medical Technology, Institute of Biotechnology in Medicine, National Yang-Ming University, Shih-Pai, 112, Taipei, Taiwan, ROC.
Wang
Y C
YC
Lin
C H
CH
Kuo
R L
RL
Liu
W T
WT
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D005434:Flow Cytometry; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006561:Herpes Simplex / Q000175:diagnosis*; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D008856:Microscopy, Fluorescence; D010957:Plasmids; D012680:Sensitivity and Specificity; D014162:Transfection; D014709:Vero Cells
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins