Health care workers and patients may acquire infections during provision and acquisition of health services, respectively, due to exposure to microorganisms in health care facilities. Studies have shown that compliance with standard infection prevention strategies is low among healthcare service providers. This study investigated the factors influencing compliance with infection prevention and control practices among health care workers,  barriers to compliance with these guidelines in Seme Sub County, Kisumu County. Data collection was done by  an observational checklist and Key Informant Interviews. Quantitative,data was subjected to content analysis. A total of 109 healthcare workers from all the health care facilities in Seme Sub-County participated in the study. About 20% of the facilities had written IPC policy guidelines. County and Subcounty hospitals and only one health center had IPC committee, while none of the dispensaries had an IPC committee in place. Majority of the HCWs reported a good uptake of IPC domains; hand washing (80.7%), waste segregation (94.5%) and injection safety (84.9%), although upon observation, only a small proportion of healthcare facilities followed the IPC domains. Majority of staff mentioned lacked of training on infection prevention and control practices, and the major barriers to compliance included lack of dissemination of guidelines, inadequate equipment and supplies, and ignorance among healthcare workers.  County level of healthcare facility was significantly associated with good uptake of handwashing (p=0.04), while nurses were more likely to have good uptake of handwashing and waste segregation compared to medical officers (p= 0.04). The findings in this study provided insight into individual and hospital related factors associated with compliance with standard precautions of infection prevention and control. These findings suggest need for improvement on infection prevention and control within the facilities. It is necessary for the hospital administration to reactivate IPC committee in all major facilities and conduct regular audits and in-service training to enhance good compliance and implementation of IPC. The ministry of health should urgently put up interventions to improve compliance with IPC in all healthcare facilities in Seme subcounty.