The epidemic characteristics of human immunodeficiency virus type 1 (HIV-1) in Zhejiang Province have not been systematically identified. This study presented a dynamic analysis of HIV-1 subtype variation in Zhejiang from 2004 to 2008, based on the surveillance of molecular epidemiology or drug resistance. CRF01_AE was the major strain (43.5 %) spreading across the province, second by B/B' (17.9 %), CRF07_BC (17.1 %), and CRF08_BC (13.7 %). The strains were mainly transmitted by heterosexual contact. Novel recombinant strains and vertical transmission were occasionally reported. Floating population from other provinces accounted for a significant number of HIV-1 cases in Zhejiang. These data may provide us rational intervention strategy for further control of HIV dissemination.