Healthcare-associated infections continue to be a major problem that endangers patient safety, especially for critically ill hospitalized patients. Reducing the risk of this infection can be achieved through improving compliance with infection control guidelines and identifying and controlling the associated factors. This study aim to investigate factors impacting compliance with infection control guidelines and increasing the risk of healthcare-associated infections in neonatal intensive care units. A qualitative approach, a one-to-one interview design, was used, between April 15 and May 15, 2021, to interview ten participants who worked in the neonatal intensive care unit at a Saudi Arabian referral hospital. Through data analysis, two major themes and four sub-themes were found, which include a factor related to the workforce (shortage of staff and lack of knowledge) and factors related to the workplace (shortage of supplies and lack of monitoring). In conclusion, healthcare-associated infections in neonatal intensive care units are a serious issue that could lead to an increase in mortality and morbidity rates among patients. Thus, healthcare settings' commitment to improve compliance with the infection control guidelines is essential to lowering the incidence of healthcare-associated infections. This can be achieved through proper staffing, sufficient supplies, and effective monitoring.