A dual reporter cell assay for identifying serotype and drug susceptibility of herpes simplex virus

Wen Wen Lu, Jun Ren Sun, Szu Sian Wu, Wan Hsuan Lin, Szu Hao Kung*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

A dual reporter cell assay (DRCA) that allows real-time detection of herpes simplex virus (HSV) infection was developed. This was achieved by stable transfection of cells with an expression cassette that contains the dual reporter genes, secreted alkaline phosphatase (SEAP) and enhanced green fluorescent protein (EGFP), under the control of an HSV early gene promoter. Baby hamster kidney (BHK) and Chinese hamster ovary (CHO) cell lines were used as parental cell lines because the former is permissive for both HSV serotypes, HSV-1 and HSV-2, whereas the latter is susceptible to infection only by HSV-2. The DRCA permitted differential detection of HSV-1 and HSV-2 by observation of EGFP-positive cells, as substantiated by screening a total of 35 samples. The BHK-based cell line is sensitive to a viral titer as low as a single plaque-forming unit with a robust assay window as measured by a chemiluminescent assay. Evaluations of the DRCA with representative acyclovir-sensitive and acyclovir-resistant HSV strains demonstrated that their drug susceptibilities were accurately determined by a 48-h format. In summary, this novel DRCA is a useful means for serotyping of HSV in real time as well as a rapid screening method for determining anti-HSV susceptibilities.

Original languageEnglish
Pages (from-to)97-104
Number of pages8
JournalAnalytical Biochemistry
Volume415
Issue number2
DOIs
StatePublished - 15 Aug 2011

Keywords

  • Antiviral susceptibility
  • Dual reporter cell assay
  • Enhanced green fluorescent protein
  • Herpes simplex virus
  • Secreted alkaline phosphatase
  • Serotyping

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