The optimization and validation of the glycoprotein ELISA assay for quantitative varicella-zoster virus (VZV) antibody detection
Article first published online: 24 OCT 2006
Copyright © 2006 Wiley-Liss, Inc.
Journal of Medical Virology
Volume 78, Issue 12, pages 1679–1687, December 2006
How to Cite
Hammond, O., Wang, Y., Green, T., Antonello, J., Kuhn, R., Motley, C., Stump, P., Rich, B., Chirmule, N. and Marchese, R. D. (2006), The optimization and validation of the glycoprotein ELISA assay for quantitative varicella-zoster virus (VZV) antibody detection. J. Med. Virol., 78: 1679–1687. doi: 10.1002/jmv.20754
- Issue published online: 24 OCT 2006
- Article first published online: 24 OCT 2006
- Manuscript Accepted: 7 AUG 2006
- quantitative assay;
Varicella is a highly contagious viral disease found throughout the world. A live-attenuated Varicella-Zoster virus (VZV) vaccine (Oka/Merck strain), VARIVAX™, was licensed in the United States (US) in 1995 and was made a part of the US recommended childhood vaccination schedule in 1996. The immune response to VZV-containing vaccines has been measured using an enzyme-linked immunosorbent assay (ELISA) to detect antibodies to glycoproteins from VZV. A correlate for protective immunity has been established between anti-VZV glycoprotein antibody levels and protection against breakthrough varicella in children, and this correlate is used as the primary immunogenicity endpoint in clinical trials with VZV-containing vaccines. The performance of the “first generation” validated version of the assay was recently reevaluated in order to identify areas for improvement. Specific format and reagent changes were implemented, with the goal of improving assay consistency by maintaining tighter control over assay processes and reagents. An extensive validation of the “second generation” gpELISA was undertaken in order to characterize the updated assay. In this article, we describe the gpELISA method, detail the procedures used to evaluate assay performance, and present the operating characteristics of the second generation gpELISA. J. Med. Virol. 78:1679–1687, 2006. © 2006 Wiley-Liss, Inc.