Objective To explore the on-the-job training mode for infection prevention and control full-time staff in basic-level hospitals, and provide reference for managers to improve on-the-job training program. Methods 109 trainees participated in the second session of professional training course for infection prevention and control full-time staff in basic-level hospitals in Suqian City were selected as research objects. Competency-oriented professional training mode with diversified training forms (including panoramic case teaching, clinical infection prevention and control manipulation skill training, clinical practice, theoretical and technical assessment before and after training, and self-assessment by adopting the core competence scale of full-time staff in healthcare-associated infection[HAI] management) supplemented to the traditional training (theoretical teaching for 2-3 days) were adopted. Results 109 trainees were from 31 township healthcare centers and 10 from community health service centers, all with full-time or part-time work experience in HAI management ≥ 6 months. After the competency-oriented training for infection prevention and control staff, the core competence, theoretical achievement and comprehensive emergency response ability of the infection prevention and control full-time staff were all higher than before training, difference was statistically different (all P < 0.001). The total score of the core competence of infection prevention and control full-time staff after training was (11.16±4.96) points higher than before training. The total satisfaction degree of the training effect of infection prevention and control staff in basic-level hospitals was 94.50% (103/109). Conclusion The competency-oriented on-the-job training mode for infection prevention and control full-time staff in basic-level hospitals achieved training according to professional requirements, implemented assessment according to professional competence, which can improve the work ability of infection prevention control full-time staff in basic-level hospitals.