Major pharmaceutical guidelines have identified Dolutegravir (DTG) as a critical medicine in the first-line treatment of HIV/AIDS. The goal of the study was to correlate DNA UGT1A1 gene expression in hyperbilirubinemia and central nervous system-related adverse reactions following oral Dolutegravir administration, using a NanoDrop-1000 spectrophotometer and rt-PCR. 52 seropositive patients were recruited using a standardized questionnaire having different components, including gender, demographic data, educational qualifications, when Highly Active-Antiretroviral Therapy (HAART) was initiated, the nature of adverse or side effects experienced, modalities used to overcome such events, and other laboratory parameters were carefully collected. Informed consent was obtained from all potential participants after an adequate explanation of the study and possible areas they would be involved, as well as samples to be collected. Blood samples were collected via venous puncture from all participants, and the same was used to quantify the level of UGT1A1 expressed in each participant using the NanoDrop-1000 spectrophotometer and rt-PCR. The outcome was compared with the side effects or adverse events experienced by each participant using ANOVA. The NanoDrop-1000 spectrophotometric results showed adequate DNA yield from 0.65 (lowest) to 104.71 ng/µl (highest) at 260nm and 280nm, respectively. At 260nm, the absorbance ranges from 0.01 to 2.09, while at 280nm from -0.01 to 1.12, respectively. Some of these participants reported having some level of anxiety, consistent headache, insomnia, drowsiness, dizziness, etc., which are clear neuropsychiatric symptoms, loss of appetite, dry throat, and anemia as adverse reactions due to HAART. There was a positive but mildly significant correlation between UGT1A1 gene expression and CNS-related adverse reactions and hyperbilirubinemia, with a p-value of 0.08.