Human T cell lymphotropic virus type 2 (HTLV-2) has been shown to be endemic in Brazilian Indians and among intravenous drug users in urban areas, but transmission of this infection seems to be infrequent in the general population living in urban areas in Brazil. Six persons in three generations of a Brazilian family were evaluated to assess HTLV-2 transmission and its molecular features in the positive cases. The index was detected during screening (HTLV EIA) of donated blood in Fundação Hemominas, Belo Horizonte, Brazil. Confirmatory serological test and viral typing were performed by Western blotting and polymerase chain reaction. The family consisted of husband, wife (index case), three daughters, and the mother of the index case. The husband and one daughter were found positive, thus pointing to horizontal and vertical transmission. The husband was a truck driver, who reported casual sex during frequent traveling. The positive daughter was breast-fed for 3 months, as opposed to the remaining two (seronegative), who breast-fed for 1 month. The index case's mother was negative. To identify HTLV-2 subtype(s), phylogenetic analysis of the noncoding long terminal repeat region and part of the env and tax coding regions was performed. These new isolates from Belo Horizonte are related to subtype IIa but present a molecular variant with extended tax, previously reported in subtype IIc. Analyzing both LTR and env regions, the family's sequences clustered with isolates of Brazilian intravenous drug users and transfusion transmitted virus.