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Human T‐lymphotropic virus type 1 (HTLV‐1) infection is an important health issue that affects a variety of endemic areas. The Khorasan province, mainly its capital Mashhad in northeastern Iran, was reported to be as one of these endemic regions. Torbat‐e Heydarieh, a large city Southwest border to Mashhad with a segregated population was investigated for the prevalence and associated risk factors of HTLV‐1 infection in 400 randomly selected individuals. Blood samples were tested for the presence of HTLV‐1 antibodies via the ELISA method and then were confirmed by an Immunoblot test. For the presence of HTLV‐1 in lymphocytes of infected subjects, PCR was performed on LTR and TAX regions. DNA sequencing of LTR fragment was also carried out to determine the phylogenetic of HTLV‐1, using the Maximum likelihood method. HTLV‐1 sero‐reactivity (sero‐prevalence) among the study population …