Japanese encephalitis virus (JEV) and other arboviruses are demonstrating an emergence in the southern part of New Guinea Island. JE was previously unknown in this part of the world until 1995 when it was found in the Torres Strait, northern Australia. In this study 96 sera collected from residents of the Timika region of Irian Jaya were tested for antibodies to JEV and related arboviruses by epitope-specific blocking ELISA. Of the 9 sera deemed to be positive for JEV antibodies by ELISA, 5 were collected from persons indigenous to Timika, and who had not travelled to regions where JE is known to be active. This indicates that these individuals were infected with JEV in the Timika area and supports a recent report of a clinical case of JE in this region. Non-immune expatriates visiting or working in the Lowland areas of Irian Jaya and/or Papua New Guinea should consider immunization against JE. Precautions should always be taken to avoid being bitten by any mosquito both in the daytime and at night.