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Background: Anxiety is common among sexual health service users. Accessible, anonymous online sexual health services may offer opportunities to connect users with mental health services, but little is known about anxiety in these settings. We sought to characterise expressions of anxiety among chat users and nurse responses to anxiety.Methods: We conducted inductive thematic analysis of transcripts from an anonymous online sexual health chat service moderated by sexual health nurses.Results: Among chat users, we identified: worry, anxiety, and emotional distress, particularly regarding HIV transmission risk, testing, and symptoms; exaggerated appraisal of HIV-transmission risk associated with sex-related shame and stigma; and patterns of anxiety that were unresolved by HIV education or testing interventions. Although nurses recognised and acknowledged anxiety, their responses to this anxiety varied; some provided anxiety management information, while others offered sexual health education and risk assessment.Conclusions: Targeted interventions addressing HIV-related stigma and anxiety among online sexual health service users are needed to facilitate connections to appropriate mental health supports. [ABSTRACT FROM AUTHOR]