ABSTRACT Women infected with Human immune deficiency virus (HIV) are five times more likely to be diagnosed with cervical cancer, because HIV infection weakens the immune system thus reducing the body's ability to fight infections that are more likely to lead to cervical cancer, Human Papilloma Virus (HPV) causes cervical cancer more in HIV positive women than those who are not infected. Therefore, the Kenya Ministry of Health requires all sexually active, HIV positive females aged above 15 years to be screened for cervical cancer. However, there is still a paucity of data on the determinants of the uptake of cervical cancer screening among HIV positive women. This study aimed to find out the determinants of uptake of cervical cancer screening among HIV positive women enrolled in the comprehensive care center (CCC) of Kisii Teaching and Referral Hospital (KTRH). A mixed retrospective cross-sectional study with both quantitative and qualitative components was conducted at the KTRH CCC in which a sample size of 255 HIV infected women were identified by systematic sampling for the primary data; 10 health workers purposively for the key informant interview (KIi), and two focus group discussions with 9 and 12 participants respectively. Quantitative data was coded and managed using SAS version 9.2 while qualitative data was coded, synthesized and grouped into exhaustive categories in line with the study objectives, then analyzed thematically. The mean age of primary respondents was 34 years. About 53% of the respondents associated cervical cancer with sexually transmitted disease (STD) while varying proportions said cervical cancer was preventable and curable, and alluded to the importance of cervical cancer screening. Stigma related to sexually transmitted diseases, fear of abnormal results and cost were perceived to affect the uptake of cervical cancer screening as only 36.08% of the women had been screened for cervical cancer which denoted low uptake of the services. Further, according to this study, HIV positive women in comprehensive care centers had inadequate knowledge of cervical cancer screening. The study recommends to the Ministry of Health (MoH) to have patient and community sensitization on cervical cancer screening, mobilize resources for HPV vaccination prevention programme and strengthen the integration of the cervical cancer screening services in CCC as programmatic interventions to improve the uptake of cervical cancer screening services by women infected with HIV at KTRH CCC.