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

      Journal of Virological Methods
      Animals, Cell Line, Costs and Cost Analysis, Evaluation Studies as Topic, Immunoassay, economics, methods, Lymphocytic choriomeningitis virus, immunology, isolation & purification, Viral Plaque Assay, Virology

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          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.

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