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Introduction: Low uptake of HIV testing and services, including pre-exposure prophylaxis (PrEP), in Thai men who have sex with men (MSM) and transgender women (TGW) may be due to the inaccuracy in self-risk assessment. This study investigated the discordance between self-perceived HIV risk and actual risk. Methods: Data were obtained between May 2015 and October 2016 from MSM and TGW enrolled in key population-led Test and Treat study in six community health centres in Thailand. Eligible participants were at least 18 years old, Thai national, had sex with men, had unprotected sex with a man in the past six months or had at least three male sex partners in the past six months, and were not known to be HIV positive. Baseline demographic behavioural characteristics questionnaires, including self-perceived HIV risk, were self-administered. Participants received HIV/STI (syphilis/gonorrhoea/chlamydia) testing at baseline. Participants who self-perceived to have low risk, but engaged in HIV-susceptible practices were categorized as having risk discordance (RD). Regression was conducted to assess factors associated with RD among MSM and TGW separately. Results: Of the 882 MSM and 406 TGW participants who perceived themselves as having low HIV risk, over 80% reported at least one of the following: tested HIV positive, engaged in condomless sex, tested positive for a sexually transmitted infection sexually transmitted infection (STI; or used amphetamine-type stimulants. Logistic regression found that living with a male partner (p = 0.005), having never tested for HIV (p = 0.045), and living in Bangkok (p = 0.01) and Chiang Mai (p < 0.001) were associated with increased risk discordance among MSM. Living with a male partner (p = 0.002), being less than 17 years old at sexual debut (p = 0.001), and having a low knowledge score about HIV transmission (p < 0.001) were associated with increased risk discordance among TGW. However, for TGW, being a sex worker decreased the chance of risk discordance (p = 0.034). Conclusions: Future HIV prevention messages need to fillin the gap between self-perceived risk and actual risk in order to help HIV-vulnerable populations understand their risk better and proactively seek HIV prevention services. Keywords: men who have sex with men; transgender women; HIV prevention; risk perception; risk discordance; HIV infection, 1 | INTRODUCTION Thailand has made remarkable progress in reducing new HIV infections over the past years [1], and its trajectory is well-set to achieve UNAIDS' 90-90-90 goals [2]. Of [...]