ABSTRACT: Background: Human parvovirus B19 (B19V) can cause anemia in some patients, including those with compromised immunity system. There are a few studies on molecular epidemiology of B19V and its association with anemia in Iran. Therefore, the aim of this study was to determine the B19V DNA, IgM, IgG, genotyping, and viral load in HIV patients in different groups of pregnant women, general population, injection drug users (IDU), and Elite controllers. Also, the possible association of B19V with anemia was studied. Methods: In this case-control study, B19V DNA, anti-B19V IgM, anti-B19V IgG, viral load, and hemoglobin level were assessed in 113 HIV positive patients and 72 healthy controls. Also, CD4+ T cell counts and HIV load were measured in the patients' group. All statistical analyses were done using STATA 14.2 software (Stata Corporation, College Station, Texas, USA). P value < 0.05 was considered statistically significant. Results: Among HIV patients, 19 (16.8%) cases had B19V DNA, 3 (2.7%) had B19V IgM, and 7 (6.2%) had B19V IgG. In control group, the prevalence of B19V DNA, IgM, and IgG was 6 (8.33%), 7(9.7%), and 19 (26.4%), respectively. In subpopulations based on transmission routes, general population had the highest B19V IgG and DNA positivity prevalence and viral load level. There was no significant association between B19V antibodies and DNA with anemia. Conclusion: The results demonstrated that B19V infection cannot be considered as a high-risk factor for anemia in adult HIV patients. However, further studies are needed to determine the exact role of B19V infection in HIV patients.