The hypothesis that human T-cell lymphotropic virus type I (HTLV-I) infection is associated with multiple sclerosis (MS) was tested by using primers specific to gag and pol regions of HTLV-I in an analysis that used polymerase chain reactions. No amplification of DNA from patients with MS was detected with primers for either region. After application of a more sensitive scheme with use of nested primers, however, half the samples, including patients with MS and normal control subjects, were found to contain pol DNA sequences. No sequences related to the HTLV-I gag region were detected among patients with MS by using nested primers. Sequences of the amplified HTLV-I pol genomes were determined. Regardless of their origin (MS or normal control), the pol region sequences were similar to HTLV-I sequences reported by other investigators. We conclude that HTLV-I does not have a distinct association with MS, but HTLV-I-related sequences, although in extremely low copy number, may be present in human genomes.