Quantification of lymphocytic choriomeningitis virus with an immunological focus assay in 24- or 96-well plates

J Virol Methods. 1991 Jun;33(1-2):191-8. doi: 10.1016/0166-0934(91)90018-u.


Titers of lymphocytic choriomeningitis virus (LCMV) were determined on adherent fibroblast cell lines in 24- or 96-well plates. After absorption of virus by cells and 48 h incubation under a methylcellulose overlay, cell monolayers were fixed with 4% formaldehyde in phosphate-buffered saline, permeabilized by incubation in 0.5% Triton X-100 in balanced salt solution and then stained with a monoclonal rat anti-LCMV and a peroxidase-labeled second stage antibody. The sensitivity of the assay is within a factor of 2-4 of conventional plaquing methods. The method also detects poorly or non-plaquing LCMV isolates, and therefore drastically reduces the need for titration of LCMV in mice. The method is quicker (2-3 days), as compared to conventional methods (4-6 days) and less expensive in terms of work and materials.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Costs and Cost Analysis
  • Evaluation Studies as Topic
  • Immunoassay / economics
  • Immunoassay / methods*
  • Lymphocytic choriomeningitis virus / immunology
  • Lymphocytic choriomeningitis virus / isolation & purification*
  • Viral Plaque Assay / economics
  • Virology / economics
  • Virology / methods*