The use of artificial intelligence (AI) in healthcare both enhances patients’ quality of life and reduces the workload of healthcare professionals.To determine the extent to which physicians, midwives, and nurses working in primary care utilize AI tools, as well as to assess their levels of AI literacy, AI-related anxiety, and attitudes toward AI-based healthcare services.This descriptive, observational research design consisted of a total of 337 participants, including 170 physicians and 167 midwives/nurses, were selected using simple random sampling. Data were collected through face-to-face interviews between July 25 and August 22, 2025. The data collection instruments included the Participant Information and AI Use Form, the Meta Artificial Intelligence Literacy Scale (MAILS), the Artificial Intelligence Anxiety Scale, and the General Attitude Toward Artificial Intelligence in Health Services Scale. Data were analyzed using SPSS 27.0 and the PROCESS macro version 4.2. Independent Samples t-Test, ANOVA, Pearson Correlation Analysis, and Mediation Analysis were used, with a statistical significance level set at p<0.05.Results revealed that the most common professional uses were for reading resources and translation, while the use of virtual assistants, automated reporting, and triage applications remained limited. The main concerns reported by participants included data privacy, lack of trust in AI, insufficient legal regulations, and integration difficulties. Overall AI literacy levels were below average, with the highest competence in using AI tools and the lowest in developing new AI applications. AI anxiety levels were moderate, with ethical concerns and fear of job loss being the most prominent.Attitudes toward AI were generally moderate, with positive attitudes being more prevalent. Physicians demonstrated higher AI literacy than midwives and nurses, while younger participants scored higher than older ones. Women exhibited higher levels of AI-related anxiety. Mediation analysis indicated that AI anxiety partially mediated the effect of AI literacy on attitudes; as AI literacy increased, anxiety levels decreased, leading to more positive attitudes toward AI use in healthcare.In conclusion, enhancing the AI awareness and literacy levels of primary healthcare professionals may reduce their concerns regarding data privacy and security, strengthen their willingness to integrate AI technologies into clinical practice.