Aim: The aim of the present study was to develop and validate a three-item oral health questionnaire for use by dietitians in an ambulatory care setting to screen human immunodeficiency virus-1-infected patients at risk of oral diseases to facilitate dental referral. Methods: The study was a questionnaire-based survey of 273 participants attending a human immunodeficiency virus clinic in Sydney, Australia. Subjects completed the oral health questionnaire and the Oral Health Impact Profile-14 contemporaneously. Results: A statistically significant correlation (rho = 0.617 (95% CI 0.54, 0.69), P < 0.0001) was found between the oral health questionnaire and the Oral Health Impact Profile-14 indicating adequate validity. Sensitivity for the three-question oral health questionnaire was found to be 84% (95% CI 76, 89) with a specificity of 55% (95% CI 46, 63). The negative predictive value was 77 % (95% CI 68, 85). A single screening question performed less well compared with overall sensitivity of the three-item oral health questionnaire. Conclusion: The present study found the three-item oral health questionnaire to be a valid and sensitive screening tool to 'trigger' for further oral health assessment and referral to dental professionals. This is a useful tool for dietitians and other health-care workers involved in the multidisciplinary preventative care of people living with human immunodeficiency virus.