Introduction Early Infant Diagnosis (EID) of HIV infection in HIV-exposed infants as early as six weeks using DNA-PCR technology is critical to improve pediatric HIV/AIDS care, and reduce child morbidity and mortality. In Wakiso district, EID service provision started in 2008 but its coverage in terms of proportion of HIV exposed infants reached is not clearly known. General objective of the study To determine the magnitude and the factors affecting uptake of early infant diagnosis services at Wakiso HCIV, Wakiso districts. Methodology This was a cross-sectional study carried out at Wakiso HCIV, Wakiso district. This study determined the prevalence of HIV among studied exposed infants and young children but also determined factors which were associated with utilization of EID for HIV infection. Data was collected using a semi-structured questionnaire from the caregivers/mothers. Results: Results were generated from 201 participants, 62% HIV exposed infants had been tested for HIV within the MOH recommended period of testing within two months of age. In this study; Age of caregivers (p=0.006), type of caregiver (p=0.003), educational status of caregivers (p=0.003), occupation of caregiver (p=0.002), family size (p=0.001) and number of children the caregiver had (p=0.002) were significantly associated with uptake of EID services on bivariate analysis. On bivariate analysis, these factors were associated with early uptake of EID services. They are: Caregivers’ knowledge of age for EID (p<0.001), knowledge of frequency of EID (p<0.001) awareness of HIV testing in infants during pregnancy (p=0.002), knowledge of benefits of EID (p=0.003), knowledge that EID can detect HIV p=0.018), knowledge of appropriate of age of EID (p<0.001); and ART status of the caregiver (p<0.05) The health facility factors on bivariate analysis associated with uptake of early infant diagnosis of HIV are: Waiting time (p<0.039); availability of EID services (p<0.014). Conclusions Uptake of EID services at Wakiso HCIV is 62% which is lower than the recommended MOH target of 80%. Recommendations The District Health Team and health workers at Wakiso HCIV should create awareness among communities on the availability and benefits of early infant testing services; strengthening option B + implementation in MCH to ensure enhanced access to ART among HIV positive pregnant women; strengthen health education during antenatal care visits to ensure that topics related to HIV testing in infants are well-articulated; strengthen family planning service provision; and advocate for improved education of the girl child in order to increase access to information

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