Quantification of lymphocytic choriomeningitis virus with an immunological focus assay in 24- or 96-well plates.
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.
Institute of Pathology, University Hospital Zürich, Switzerland.
M
M
Battegay
S
S
Cooper
A
A
Althage
J
J
Bänziger
H
H
Hengartner
R M
RM
Zinkernagel
D000818:Animals; D002460:Cell Line; D003365:Costs and Cost Analysis; D005069:Evaluation Studies as Topic; D007118:Immunoassay / Q000191:economics / Q000379:methods*; D008217:Lymphocytic choriomeningitis virus / Q000276:immunology / Q000302:isolation & purification*; D010948:Viral Plaque Assay / Q000191:economics; D014773:Virology / Q000191:economics / Q000379:methods*