This study determines the validity and reliability of the Malay version of the World Health Organization Quality of Life (WHOQOL) assessment instrument in patients with human immunodeficiency virus (HIV) infection. A cross-sectional study on 157 patients with HIV seen at the Infectious Disease Unit, Hospital Raja Perempuan Zainab II, Kota Bharu, Kelantan was conducted. Factor analysis identified five major domains: physical needs, spirituality, social relationship, psychological, and environment. Significant correlation was found between each domain scores and the general health questions. The instrument was able to discriminate between asymptomatic and symptomatic HIV positive patients for all domain scores except for the spirituality domain. The internal consistency of the five domains ranged from 0.70 to 0.83. The intraclass correlation coefficient (ICC) ranged from 0.60 to 0.87 across all domains. In conclusion, the Malay version of WHOQOL-HIV BREF is a valid and reliable instrument in assessing quality of life in HIV positive patients.