Development of a Novel Efficient Fluorescence-Based Plaque Reduction Microneutralization Assay for Measles Virus Immunity
- 1 July 2008
- journal article
- research article
- Published by American Society for Microbiology in Clinical and Vaccine Immunology
- Vol. 15 (7), 1054-1059
- https://doi.org/10.1128/cvi.00008-08
Abstract
The measurement of functional measles virus-specific neutralizing antibodies is of considerable interest for vaccine-related research. In this study, we developed and standardized a simple, rapid, highly sensitive, and reproducible fluorescence-based plaque reduction microneutralization (PRMN) assay with visual and automated readout, using a recombinant measles virus engineered to express enhanced green fluorescent protein. The assay is performed in micro format, requires less time to complete (2 versus 4 to 7 days), and is less labor-intensive and less costly than the classical plaque reduction neutralization (PRN) test, widely accepted as the “gold standard” in measles serology. Two available WHO international anti-measles virus standards and one in-house reference serum were used to develop and standardize the new assay. The mean PRMN values from repeated assays were found to be similar to those reported in the literature or assigned to the WHO standards by the classical PRN assay. For validation, we used three groups of low, moderate, and high measles virus vaccine responders’ sera with moderate values of correlation in antibody levels (mIU/ml) between PRMN and the Dade Behring immunoglobulin G enzyme immunoassay (EIA). The PRMN assay was more sensitive at low antibody levels and more informative in terms of protection than this commercial EIA. In conclusion, we have developed and validated a sensitive and high-throughput measles virus-specific PRMN that can be readily used in large population-based measles studies.Keywords
This publication has 17 references indexed in Scilit:
- Plaque reduction neutralization test for measles antibodies: Description of a standardised laboratory method for use in immunogenicity studies of aerosol vaccinationVaccine, 2007
- Variations in measles vaccine–specific humoral immunity by polymorphisms in SLAM and CD46 measles virus receptorsJournal of Allergy and Clinical Immunology, 2007
- Development of a new neutralization test for measles virusJournal of Virological Methods, 2007
- Global measles eliminationNature Reviews Microbiology, 2006
- Measles immunity testing: Comparison of two measles IgG ELISAs with plaque reduction neutralisation assayJournal of Virological Methods, 2005
- The contribution of HLA class I antigens in immune status following two doses of rubella vaccinationHuman Immunology, 2004
- Selectively Receptor-Blind Measles Viruses: Identification of Residues Necessary for SLAM- or CD46-Induced Fusion and Their Localization on a New Hemagglutinin Structural ModelJournal of Virology, 2004
- Development of a Novel Vaccinia‐Neutralization Assay Based on Reporter‐Gene ExpressionThe Journal of Infectious Diseases, 2003
- Neutralizing B Cell Response in MeaslesViral Immunology, 2002
- Measles Antibody: Reevaluation of Protective TitersThe Journal of Infectious Diseases, 1990