A refined complement-enhanced neutralization test for detecting antibodies to Junin virus

A refined, complement-enhanced, plaque-reduction neutralization test was developed for measuring neutralizing antibodies against Junin (Argentine hemorrhagic fever) virus. The assay measured neutralizing antibodies after natural as well as vaccine-induced Junin virus infections. Among vaccinated individuals, titers were 2–4-fold higher than those obtained with conventional assays, without loss of specificity. Enhanced sensitivity was achieved by using a standardized complement source (vs human or animal serum) for virus dilution, incubation of virus-serum mixtures at 36°C for 2 h (vs overnight at 4°C) prior to plaque assay, control of age and density of cell monolayers, and variation in overlay conditions.