The clinical significance of human T-cell lymphotropic virus type II (HTLV-II) infection, unlike that of HTLV-I, is unknown, and the major known association of HTLV-II seropositivity is with intravenous drug abuse. Screening of blood donors for HTLV-I, now routine in North America, does not distinguish this retrovirus from HTLV-II. To find out more about the seroepidemiology of and risk factors for HTLV I and II, blood from 480,000 volunteer donors in five geographically separate US urban centres was tested for antibodies to HTLV-I/II and HIV-1. Confirmed HTLV-I/II seropositive donors were then followed up by DNA amplification to distinguish type I from type II and by interviews focusing on possible risk factors. HTLV seroprevalence was 3.3 times greater than that for HIV-1 (0.043% vs 0.013%). DNA amplification on 65 of the 207 HTLV-I/II seropositive donors revealed that 34 (52%) had HTLV-II infection and 28 (43% had HTLV-I; 3 samples were uninformative. Interviews of 49 donors showed that whereas HTLV-I was principally associated with donor origin from endemic regions, the major risk factor for HTLV-II infection was intravenous drug use. The surprisingly high rate of HTLV-II infection in US blood donors raises important public health and donor counselling issues since HTLV-I infection is associated with adult T-cell leukaemia and a neurological disorder while the pathogenicity of HTLV-II is as yet unclear.