BACKGROUND: Although there is a growing interest in the epidemiology of sedentary behaviours, it is unknown whether sedentary behaviour questionnaires are broad markers of sedentary time. The aims of this study were to determine the: (i) reliability of the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) screen time-based sedentary behaviour questionnaire; and (ii) its validity, i.e. the ability of the questionnaire to correctly rank adolescents according to the objectively measured sedentary time. METHODS: A total of 183 adolescents (104 females aged 12.5-17.5 years) were involved in the reliability study. Participants completed the HELENA sedentary questionnaire twice (1 week apart). The validity study comprised 2048 (1212 females) adolescents (12.5-17.5 years of age) included in the HELENA cross-sectional study. Questions included television viewing, computer games, console games, Internet for study and non-study reasons and study during week and weekend days. We compared median values of sedentary time, using accelerometers, by tertiles of self-reported sedentary behaviours and their sum (composite sedentary score). RESULTS: Reliability study: κ-values showed a good agreement (>0.7), except for Internet for study reasons (0.46 weekdays, 0.33 weekend). The questionnaire correctly classified boys' sedentary time when analysed by specific behaviours and by a composite sedentary score. In girls, median values of objectively measured sedentary time were not different across tertiles of self-reported sedentary behaviours or the composite sedentary score. CONCLUSION: The HELENA sedentary questionnaire is reliable, yet only correctly classifies objectively measured sedentary time in boys.