This paper aimed to investigate the brain activity of human immunodeficiency virus (HIV) positive patients with normal cognition during unilateral hand movement and whether highly active antiretroviral therapy (HAART) could affect the brain function. Functional magnetic resonance imaging (fMRI) was performed for 60 HIV positive (HIV+) subjects and −42 healthy age-matched right-handed control subjects. Each subject was evaluated by the neuropsychological test and examined with fMRI during left and right hand movement tasks. HIV+ subjects showed greater activation in anterior cingulum, precuneus, occipital lobes, ipsilateral postcentral gyrus and contralateral cerebellum compared with control group during right hand movement task. However, during left hand movement no statistically significant difference was detected between these two groups. HAART medication for HIV+ subjects lowered the increased activity to normal level. Meanwhile patients receiving the regimen of zidovudine, lamivudine and efavirenz showed lower activity at bilateral caudate and ipsilateral inferior frontal gyrus in comparison with subjects receiving other HAART regimens. Therefore, HIV+ subjects demonstrated brain asymmetry in motor cortex, with increased activity present during right hand movement but absent during left hand movement. HAART proves effective in HIV+ subjects even with normal cognition and the specific regimen of HAART could prevent cerebral abnormal functions. Meanwhile, this study validates that during motor tasks, fMRI can detect the brain signal changes prior to the occurrences of other HIV- associated dysfunctions.