title: An improved respiratory syncytial virus neutralization assay based on the detection of green fluorescent protein expression and automated plaque counting.
abstract: BACKGROUND
  Virus neutralizing antibodies against respiratory syncytial virus (RSV) are considered important correlates of protection for vaccine evaluation. The established plaque reduction assay is time consuming, labor intensive and highly variable.
  
  
  METHODS
  Here, a neutralization assay based on a modified RSV strain expressing the green fluorescent protein in combination with automated detection and quantification of plaques is described.
  
  
  RESULTS
  The fluorescence plaque reduction assay in microplate format requires only two days to complete and is simple and reproducible. A good correlation between visual and automated counting methods to determine RSV neutralizing serum antibody titers was observed.
  
  
  CONCLUSIONS
  The developed virus neutralization assay is suitable for high-throughput testing and can be used for both animal studies and (large scale) vaccine clinical trials.
authors.affiliation: Department of Vaccine Research, Vaccinology, National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: van Remmerden
authors.forename: Fang
authors.initials: F
authors.lastname: Xu
authors.forename: Mandy
authors.initials: M
authors.lastname: van Eldik
authors.forename: Jacco G M
authors.initials: JG
authors.lastname: Heldens
authors.forename: Willem
authors.initials: W
authors.lastname: Huisman
authors.forename: Myra N
authors.initials: MN
authors.lastname: Widjojoatmodjo
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood* / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D002460:Cell Line; D002522:Chlorocebus aethiops; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D025202:Molecular Diagnostic Techniques; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000379:methods*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000276:immunology*; D012136:Respiratory Syncytial Viruses / Q000235:genetics / Q000276:immunology*; D014709:Vero Cells; D010948:Viral Plaque Assay
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D049452:Green Fluorescent Proteins