Antigenic relationships between attenuated and pathogenic strains of Junin virus (JV) were investigated. Five strains of either human or rodent origin were tested by cross-neutralization assay with hyperimmune antisera, raised in rabbits, against each strain. Polyclonal antisera could be used to distinguish among these JV strains, as the titer values differed significantly with ratios of homologous to heterologous titers, which ranged from 1.3 to 22.3. This demonstrates, independent of their virulence, a heterogeneity among the JV strains tested. The relatedness among JV strains was expressed quantitatively through a dendrogram based on taxonomic distance coefficients. The field strains of JV were grouped into two clusters, according to their geographic origin.