Viruses with genomes consisting of 12 segments of double-stranded (ds) RNA were isolated from mosquito pools collected in Indonesia in 1980 and 1981. The genome segments of 11 virus isolates exhibited two distinct electrophoretic patterns in agarose gels, and the viruses probably represent distinct groups of Coltiviruses. Ten of the 11 virus isolates produced recognizably similar PAGE profiles of two groups of 6 segments (6-6) with 3 distinct profile variants, and the remaining virus isolate displayed a unique 6-5-1 profile. These profiles were distinct from that of Colorado tick fever (CTF) virus RNA. The genetic relatedness of the isolates was examined by RNA-RNA hybridization. The viruses with a 6-6 profile were closely related in the majority of their segments, with segments 7, 11, and 12 being variant; and segment 9 the most divergent. The segments of the virus with the unique PAGE profile (6-5-1) did not cross-hybridize with any of the other viruses. Similarly, RNA from virus isolates from Indonesian did not cross-hybridize with the segments of CTF virus. These data indicate that two additional genotypes are present in the Coltivirus genus.