Objective: To investigate the validity of hearing screening with hearTest smartphone-based audiometry and to specify test duration addressing the two response modes and hearing loss criteria.Design: A diagnostic accuracy study comparing hearing screening with conventional audiometry.Study sample: Three hundred and forty individuals, aged between 5-92 years.Results: Of the 340 participants, 301 undertook all test procedures (273 adults and 28 children). Sensitivity and specificity were >90% for hearTest hearing screening to identify disabling hearing loss for both response modes with adults and children. We found similar sensitivity in identifying any level of hearing loss for both response modes in children, with specificity >80%, and for the self-test mode in adults. Low specificity was observed when identifying any level of hearing loss in adults using the test-operator mode. In adults, there was a significant difference between test duration for the test-operator and self-test modes.Conclusion: Hearing screening using hearTest smartphone-based audiometry is accurate for the identification of both disabling hearing loss and any level of hearing loss in adults and children in the self-test response mode. The test-operator mode is also an option for children; however, it does not provide good accuracy in identifying mild level of hearing loss in adults.