Herpes Simplex Virus Type 1 Propagation, Titration and Single-step Growth Curves

Linda Grosche; Katinka Döhner; Alexandra Düthorn; Ana Hickford-Martinez; Alexander Steinkasserer; Beate Sodeik

doi:10.21769/BioProtoc.3441

Herpes Simplex Virus Type 1 Propagation, Titration and Single-step Growth Curves

Bio Protoc. 2019 Dec 5;9(23):e3441. doi: 10.21769/BioProtoc.3441.

Authors

Linda Grosche¹, Katinka Döhner², Alexandra Düthorn¹, Ana Hickford-Martinez², Alexander Steinkasserer¹, Beate Sodeik²

Affiliations

¹ Department of Immune Modulation, Universitätsklinikum Erlangen, Erlangen, Germany.
² Institute of Virology, OE5230, Hannover Medical School, Hannover, Germany.

Abstract

Given the endemic seroprevalence of herpes simplex viruses (HSV), its associated human diseases, and the emergence of acyclovir-resistant strains, there is a continuous need for better antiviral therapies. Towards this aim, identifying mechanistic details of how HSV-1 manipulates infected cells, how it modulates the immune responses, and how it causes diseases are essential. Measuring titers and growth kinetics of clinical isolates and viral mutants are important for a thorough characterization of viral phenotypes in vitro and in vivo. We provide protocols for the preparation as well as titration of HSV-1 stocks, and explain how to perform single-step growth curves to characterize the functions of viral proteins or host factors during infection. In particular, we describe methods to prepare and characterize high-titer HSV-1 stocks with low genome to titer ratios that are required for infection studies in cell culture and animal experiments.

Keywords: Herpes simplex virus type 1; Plaque assay; Single-step growth curve; Virus propagation.