Focus luminescence assay: macroscopically visualized foci of human cytomegalovirus and varicella zoster virus infection

Abstract The plaques or foci of certain viruses due to their small size have to be counted microscopically e.g., human cytomegalovirus (HCMV). The focus luminescence assay (FLA) described below generates macroscopic images as a result of the magnification due to scattered emitted light, and provides a hard copy using autoradiography or video imaging. Foci are detected according to an immunohistochemical protocol with horseradish peroxidase or alkaline phosphatase antibody conjugates which convert substrate into a luminescent product. Detection of varicella zoster virus (VZV) foci developed with a specific substrate-enhancer combination was so sensitive that 20-times lower primary antibody concentrations were effective than those required for conventional immunohistochemical staining. This method for HCMV and VZV may allow quantitative infectivity and focus reduction assays for viruses which produce little or no CPE.