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Objectives Doxycycline post-exposure prophylaxis (doxyPEP) has been shown to reduce syphilis and chlamydia incidence among gay, bisexual and other men who have sex with men (gbMSM) and transgender women (TGW). However, evidence among other populations remains limited. This study aimed to assess knowledge, attitudes, willingness and use of doxyPEP among sexual health clinic attendees and healthcare workers in Dublin.Methods Data were collected via two anonymous surveys in two sexual health services for service users and healthcare workers. Surveys were collected between January and March 2024. Logistic regression analyses explored factors associated with willingness to use doxyPEP.Results Among 378 clinic attendees (median age 32), 72% (272/378) identified as gbMSM, TGW, non-binary or gender fluid; 17% (64/378) cisgender women and 11% (42/378) heterosexual men. Overall, 62% (235/378) expressed willingness to use doxyPEP, with higher willingness observed in the gbMSM, TGW, non-binary and gender fluid group (70%, 190/272). This group also reported the highest level of knowledge of doxyPEP compared with other groups: 107/272 (39.3%). After adjustment, logistic regression analyses revealed that individuals with prior sexually transmitted infection (STI) diagnoses were more willing to use doxyPEP (adjusted OR (aOR) per number of STI diagnosed 2.00, 95% CI 1.28 to 3.03, p=0.002). 9% (35/378) reported doxyPEP use in the past 1 year. Among 52 healthcare workers, 81% (42/52) were aware of doxyPEP studies, 62% (32/52) believed it could reduce STIs, but 89% (46/52) had concerns, primarily around antimicrobial resistance.Conclusions This is the first study evaluating doxyPEP attitudes in Ireland. Willingness to use doxyPEP was high among individuals at greatest STI risk, supporting its potential for targeted implementation. There was also interest among cisgender women and heterosexual men. However, clinician concerns about antimicrobial resistance remain a barrier. These findings highlight the need for national guidelines, ongoing surveillance and research, including among heterosexual men and cisgender women, to inform equitable and safe doxyPEP prescription.