ObjectiveTo evaluate the effectiveness of Kirkpatrick fourlevel training evaluation model on evaluating healthcareassociated  infection(HAI) training program,so as to improve the training quality of HAI  knowledge gradually.MethodsThrough combination of questionnaires, field survey and expert interviews, the effectiveness of HAI knowledge training among new employees was  evaluated by Kirkpatrick fourlevel training evaluation model.ResultsA total of 192 new employees participated in the training program, including 132 (68.75%) doctors and technicians (D&T), 60 (31.25%) nurses, the response rate was 100%, the overall satisfaction rate was 89.41%. The posttraining score was significantly increased  compared with pretraining score(［83.43±5.56］ vs ［54.22±5.13］,t=54.15, P<0.001); The yearend assessment score was significantly different compared with posttraining score (［82.38±4.28］ vs ［83.43±5.56］, t=2.16,P=0.03), the yearend assessment score decreased by 1.05 points compared with posttraining score. Multiple comparison of pre and posttraining score on HAI knowledge among D&T and nurses were conducted, except the scores on “hospital cleaning, disinfection and sterilization and isolation” as well as “aseptic technique”, the scores on the other aspects were significantly different (P<0.05). Multiple comparison of posttraining and yearend assessment score on HAI knowledge among D&T and nurses showed that the scores on most  knowledge points have declined (P<0.05). Doctors and technicians’ knowledge on “hand hygiene” and “occupational safety” decreased,  and the nurses’ knowledge on “bacterial resistance mechanisms” and “rational use of antimicrobial agents” decreased.ConclusionKirkpatrick fourlevel training evaluation model can objectively reflect the internal and external effectiveness of training, the training has achieved a significant shortterm effect, but  longterm effect is not obvious. The quality of training can be improved by perfecting training mode and establishing a systematic evaluation system.