In the United States, HIV has evolved from an acute disease to a chronic illness making health-related quality of life a pre-eminent goal for many persons living with HIV (PLWH). There have been a number of HIV-specific quality-of-life instruments developed, but little attention has been paid to the validation of standardized nondisease-specific quality-of-life instruments tailored to PLWH. The goal of this research was to validate the Patient-Reported Outcomes Measurement Information System (PROMIS)-29, a questionnaire that measures health-related quality of life in PLWH. A sample of 1306 PLWH completed an online anonymous survey assessing their symptom experience and health-related quality of life. A subsample of 209 participants completed another questionnaire 30 days later. The subscales of the PROMIS-29 showed high internal consistency reliability (range = 0.87-0.97). The PROMIS-29 detected differences in health-related quality of life in those persons who reported an AIDS diagnosis compared to those who did not report an AIDS diagnosis. The PROMIS-29 has demonstrated reliability, validity, and reproducibility for use in measuring health-related quality of life in PLWH.