Health care associated infections (HCAI) are one of the most serious and complex worldwide health problems. Commitment to standard precautions (SPs) and infection control (IC) measures are important to all healthcare providers (HCP) to prevent occupational exposure to hazardous materials. Healthcare workers (HCWs) are particularly more prone to blood-borne pathogens through contact with infected blood and body fluids during their work. It is based on the presumption that every person is infected or colonized with an organism that could be transmitted in the healthcare setting and thus health care workers need to apply infection control practices during the delivery of health care. Health care worker at risk of health associated infection (HAIs) due to their exposure to the patients during carrying out their duties. Health care workers in primary health care centers are the first to detect and to prevent the spread of infection. Standard precautions are regarded as an effective means for protecting health care worker, patients and community. Aim of the study: The study aimed to assess the level of Knowledge and practices of infection control standard among health care workers at the primary healthcare level in Makkah City . Method: A cross sectional study enrolled HCWs (doctors, nurses, lab workers) from 10 primary healthcare (PHC) centers in in Makkah, during the April to June, 2021, Our total participants were (200). Results: the knowledge regarding standard precautions of infection control study results show the majority of participant had average information were(55.0%) while weak knowledge were(21.0%) the data ranged from(5-18) by mean ±SD(12.15±3.11), also the Nationality is a significant relation between knowledge and Practices increase in the non-Saudi healthcare workers were mean+ SD respectively (12.980±2.606, 6.603±1.510), compared to the Saudis had adequate level of knowledge Practices where (T=-5.33, T=-6.152) and P-value=<0.001. Conclusion: There was a gap between the actual and desired Knowledge and practices of Healthcare providers regarding IC. Continuing education programs are needed to improve their Knowledge and practices scores towards SPs and IC measures in order to reduce HCAI, inadequate knowledge particularly concerning the disposal of sharp instruments and hand hygiene were also detected. The studies highlighted the necessity of the provision of a comprehensive training program to ensure compliance with infection control measures by HCWs.