Antiretroviral drugs help reduce the viral load in HIV patients and prevent further progression of the virus. Hepatotoxicity has been reported among HIV patients receiving Highly Active Antiretroviral Therapy (HAART). There is therefore the need to monitor liver enzyme activity in HIV seropositive patients on different antiretroviral (ARV) regimens. This study thus aimed at determining the variation of serum transaminases; aspartate aminotransferase (AST) and alanine aminotransferase (ALT) amongst HIV/AIDS patients on different ARV regimens at the Saint Elizabeth Hospital, Shisong with respect to different ARV regimens, age groups, gender, and duration on ARV therapy. The results will guide health personnel in administering anti-retroviral therapy to HIV/AIDS patients and bring to focus the need for a liver function test before placing a patient on antiretroviral therapy. In this crosssectional exploratory hospital and laboratory-based study involving 57 participants, venous blood was collected and the absorbance and concentration of serum transaminases for each subject read using a spectrophotometer. Data obtained were analyzed using SPSS and the Chi-Square test used to determine association and significance at Pvalue ≤ 0.05. Results showed there was significant elevation of transaminases among study participants. Elevation of transaminases was more pronounced for AST (47.4%) than ALT (19.3%). Regarding the different ARV regimens, participants on a combination of Nucleoside Reverse Transcriptase Inhibitor and Non-Nucleoside Reverse Transcriptase Inhibitor (NRTI+NNRTI) had higher transaminase elevations than those who were on single or triple combinations. With respect to sex, elevation of AST levels was significantly higher (RR=1.0962) in females (48.7%) than males (44.4%). For ALT, males tend to have more elevated levels than females but the difference was not significant (RR=0.8077). There was no significant difference in transaminase levels with age groups (P>0.05).