title: The optimization and validation of the glycoprotein ELISA assay for quantitative varicella-zoster virus (VZV) antibody detection.
abstract: Varicella is a highly contagious viral disease found throughout the world. A live-attenuated Varicella-Zoster virus (VZV) vaccine (Oka/Merck strain), VARIVAXtrade mark, 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.
authors.affiliation: Vaccine and Biologics Research, Merck Research Laboratories, Wayne, Pennsylvania, USA.
authors.forename: Olivia
authors.initials: O
authors.lastname: Hammond
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.forename: Tina
authors.initials: T
authors.lastname: Green
authors.forename: Joseph
authors.initials: J
authors.lastname: Antonello
authors.forename: Robert
authors.initials: R
authors.lastname: Kuhn
authors.forename: Clifford
authors.initials: C
authors.lastname: Motley
authors.forename: Philip
authors.initials: P
authors.lastname: Stump
authors.forename: Beverly
authors.initials: B
authors.lastname: Rich
authors.forename: Narendra
authors.initials: N
authors.lastname: Chirmule
authors.forename: Rocio D
authors.initials: RD
authors.lastname: Marchese
mesh_terms: D000328:Adult; D000914:Antibodies, Viral / Q000097:blood*; D002644:Chickenpox / Q000276:immunology / Q000517:prevention & control; D019433:Chickenpox Vaccine / Q000008:administration & dosage / Q000276:immunology*; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006023:Glycoproteins / Q000276:immunology*; D006562:Herpes Zoster / Q000276:immunology / Q000517:prevention & control; D014645:Herpesvirus 3, Human / Q000276:immunology*; D006801:Humans; D007223:Infant; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D019433:Chickenpox Vaccine; D006023:Glycoproteins; D011933:Reagent Kits, Diagnostic; D014759:Viral Envelope Proteins