Background: This study aimed to evaluate the feasibility of using the HIV Self-Test in high-risk Iranian groups (MSM, FSWs, and TG people).Methods: This study was conducted in 2 main steps (the pre and post HIVST intervention and the qualitative method) between 1 October 2020 and 22 June 2021 in Tehran, Iran. The sample size needed for this study generally was 1000 people, including FSWs, MSM, and transgender individuals. The sampling method in this study was convenience sampling, while snowball sampling was used to complete the number of samples.Results: A total of 930 eligible respondents were enrolled in the study, of whom 456 (49%) were female, 49 (5.3%) were transgender (98% of TGs were male to female). The median age was 33.63 ±10.54 SD. The feasibility of the use of HIV self-tests in Iranian high-risk groups was significantly high. The majority of participants (97%) did not have any confidentiality problems while preparing or performing the test. In general, feasibility was assessed based on five questions. The overall feasibility score was 6.33 (0.824 SD). Taking tests, reading HIV test results, finding a safe place to do the test, and accessing HIV self-testing showed a high average. Conclusion: HIV self-testing was highly acceptable and feasible to permit routine HIV testing between high-risk populations efficiently.