This study aims at estimating the prevalence of Leishmania infection among HIV-infected patients through the use of non-invasive tests. The study was conducted in three Infectious Diseases Services in two large Brazilian cities, both endemic areas for visceral leishmaniasis. Three hundred and eighty-one asymptomatic patients were enrolled whose ages ranged from 19 to 58 years old; 63.5% were men; mean TCD4+ was 380 cells/μl; and mean viral load was 153800 copies/ml. All individuals were tested for Leishmania infection through: ELISA using crude Leishmania infantum (ELISA), ELISA using the recombinant K39 antigen (rK39), indirect fluorescent antibody test (IFAT) and PCR targeted to kDNA region. The tests' positivity were: 10.8% (ELISA), 3.9% (IFAT), 0.8% (rK39), 6.3% PCR and 20.2% (overall, at least one positive test), with no statistical correlation between positivity and clinical and laboratorial variables. Concordance among tests was low (Kappa <0.20). Prevalence of Leishmania asymptomatic infection was high in this population, reinforcing the need for attention in the evaluation of HIV patients from endemic areas. New efforts are needed to develop more specific and sensitive tests to diagnose Leishmania asymptomatic infection. Highly active antiretroviral therapy (HAART) seems to have a protective role against disease progression in co-infected individuals.