An application of tetrazolium (MTT) colorimetric assay for the screening of anti-herpes simplex virus compounds

Abstract

A rapid and sensitive procedure was developed for in vitro evaluation of anti-herpes simplex virus (HSV) agents. The procedure is based on spectrophotometrical assessment for viability of virus- and mock-infected cells via in situ reduction of a tetrazolium dye MTT, which has already been used for the detection of anti-human immunodeficiency virus (HIV) agents (Pauwels et al., 1988). Monolayer cells such as human embryonic fibroblast, VERO, or HeLa cells were not suitable for this purpose. Among the non-adherent cell lines examined for susceptibility to HSV type 1 (HSV-1), a B-lymphoblastoid cell line NC-37 was found to be the most sensitive. The cell line was found to have a good correlation between the viable cell number and the reduction of MTT. In addition, centrifugation of the virus-infected cells resulted in further increase of the sensitivity of NC-37 cells to HSV-1. After optimization, the method proved to be as sensitive as plaque reduction. The system simplifies significantly the assay procedures and thus permits the evaluation of larger numbers of compounds for anti-HSV-1 activity.