During the past few decades several newly recognized rodent-borne arenaviruses have been shown to be associated with severe hemorrhagic fever cases in South America. Changes in ecology and farming practices throughout the region have increased the concern over the potential public health threat posed by such emerging virus diseases. Oliveros (OLV) virus is a recently discovered arenavirus of the rodent Bolomys obscurus in Argentina. Genetic analysis of the small genomic RNA segment, which encodes the nucleocapsid protein and the envelope glycoproteins, shows that Oliveros is a novel, phylogenetically distinct member of the Arenaviridae family which differs in nucleotide sequence from the previously characterized members by approximately 35% or more. Despite this level of diversity, OLV virus possesses the same ambisense genome structure and many overall RNA and protein features in common with other arenaviruses. These data represent an important first step in the development of specific immunological and PCR diagnostic reagents to allow assessment of the prevalence and disease potential of this virus.