Isolation and partial sequencing of human T-lymphotropic virus type I (HTLV-I) variants from inhabitants of Papua New Guinea (PNG) and the Solomon Islands has confirmed the existence of virus infection in Melanesian populations. To determine the geographical distribution of seropositivity to HTLV-I in PNG we have tested 2907 serum and plasma samples collected between 1972 to 1991 from 16 different population groups. Samples were screened using a particle agglutination assay and confirmed by p21e-enhanced Western immunoblot (WB). From a total of 94 screen positive samples run on WB, 56 (60%) were confirmed positive (positive for both env and gag products) and 38 (40%) were WB-indeterminate (gag products only). The prevalence of WB-confirmed antibodies to HTLV-I in lowland and island populations ranged from 0 to 5.4%. There were no confirmed antibody positives in the highland populations surveyed. Geographically isolated populations living on the fringes of the highlands ranged in seropositivity from 0.2 to 5.8%. Two of the subjects surveyed gave WB antibody patterns characteristic of HTLV-II rather than HTLV-I infection.