Background Chicago’s HIV epidemic disproportionally affects people of color. Almost a quarter (23%) of these infections occur in Hispanics. It is important to understand sexual behaviors and HIV risk in Latino MSM and transgender women (TGW) to create targeted culturally sensitive harm reduction interventions. However, participation of minority MSM and TGW in survey-based studies is low. The main objective of the study was to understand the sexual health and of Latino MSM and TGW residing in Chicago, Illinois, United States. We herein report subject’s attitudes towards participating in the study and qualitative observations about perceived barriers to enrollment of this population. Methods This study was a cross sectional analysis of a behavioral/HIV seroprevalence survey administered during 2017-2020 to presumed HIV negative, Latino identifying, MSM and TGW individuals. The survey included questions on sexual risk, HIV knowledge and depression scores. We categorized recurrent themes of the most common reasons participants provided for declining to participate in the study. We generated descriptive statistics. Results A total of 48 community organizations assisted with recruitment. Of 149 participants screened, only 18 (12%) agreed to complete the survey. Among those who declined to complete the survey (n=131), the most common reasons given were: participants were uncomfortable answering some questions (n=59, 45.0%), participant’s did not have the time to initiate or complete the survey (n=41, 31.3%) and survey was deemed long (n=16, 12.2%). We categorized barriers to successful recruitment into the following general themes: 1) Participants were unwilling to discuss their sexual history; 2) Participant’s felt uncomfortable taking a rapid oral HIV test; 3) Participant’s lacked transportation; and 3) Participants didn’t have time to complete the survey or thought it was too long. Conclusion Despite extensive community networking, we found barriers to recruitment of high-risk Latino MSM and TGW into an HIV seroprevalence study. Further research is needed to better understand and address these barriers, and thus, increase representation of this key population in prevention studies. Disclosures All Authors: No reported disclosures