Background Although French Guiana is highly endemic for HIV-associated histoplasmosis, data from the Guiana shield are lacking. This study estimated the prevalence of histoplasmosis among PLHIV in Suriname and French Guiana using antigen assays not routinely available locally. Methods We conducted a prospective cross-sectional study (2013-2015) in the main hospitals of both countries. Consecutive adult PLHIV with fever, altered general condition, or symptoms suggestive of infection were included. In addition to standard investigations, three antigen tests were performed: polyclonal EIA on urine and serum, and monoclonal EIA on urine. Proven or probable histoplasmosis was defined using EORTC/MSGERC criteria. Results Among 478 patients (307 64% in Suriname; median CD4+ count of 114/µL), histoplasmosis was the most frequent opportunistic infection (111 patients, 23.2%). Antigen detection increased diagnosis by 208% overall (Suriname: 420%; French Guiana: 57%). Prevalence remained substantial (11.1%) in patients with CD4+ counts between 100 and 349/µL. Serum antigen testing showed the best diagnostic performance (AUC: 0.87). Thirty-day mortality was higher in patients with histoplasmosis (17.5% vs 8.4%). High antigen levels were associated with mortality. Conclusions Histoplasmosis is highly prevalent among hospitalized PLHIV on the Guiana shield. Antigen detection improves diagnosis and may help identify patients at higher risk of poor outcomes.