The primary aim of this study was to estimate HIV incidence within a high-risk population in Ho Chi Minh City (HCMC), Vietnam using both cross-sectional and prospective methodologies. A secondary aim was to develop a local correction factor for the BED and avidity index incidence assays. The research study design consisted of three phases: (1) cross-sectional, (2) prospective, and (3) BED false recent (BED FR). A total of 1619 high-risk, sexually active individuals were enrolled in the cross-sectional phase and 355 of the opiate-negative, HIV-negative women were subsequently enrolled in the prospective phase. Four-hundred and three men and women with known HIV infection duration of greater than 12 months were enrolled in the BED FR phase. The HIV prevalence for all participants in the cross-sectional phase was 15.8%. HIV incidence in the cross-sectional group was estimated using the BED IgG capture assay and AxSYM avidity index assay for recent HIV infection and incidence within the prospective cohort was determined by observations of HIV seroconversion. HIV incidence in opiate-negative women was estimated using the BED assay to be 0.8% unadjusted and 0.5% after applying the locally derived BED false recent rate of 1.7%; no seroconversions were observed in the prospective cohort. We also screened the cross-sectional samples for evidence of acute infection using nucleic acid testing, 4th generation HIV EIA, and SMARTube coupled with Genscreen and Determine diagnostic tests; no confirmed acute infections were identified by any method. HIV incidence within this opiate-negative study population was low and incidence estimates from the two methods compared favorably with each other. Incidence estimates and false recent rates using the AxSYM assay were higher: AI FRR of 2.7% and adjusted incidence of 1.7% per year (95% CI, 0.6, 2.8). By comparison, both HIV prevalence and incidence estimates for the opiate-positive group were higher.