Objective To understand the current situation and influencing factors of knowledge, attitudes, and practice (KAP) among nurses regarding disinfection after intermittent medication administration through needle-free connectors (NFC), so as to provide a basis for improving the aseptic management of NFC.Methods Convenience sampling was used to select 978 nurses from 10 hospitals in southern, central, and northern Jiangsu Province.A self-designed questionnaire was used to investigate KAP among nurses regarding disinfection of NFC after intermittent administration via NFC.Results The scores for knowledge, attitude, and practice regarding disinfection after intermittent medication administration through NFC in nurses were 10.00（5.00,20.00）, 12.00（12.00,16.00）, and 6.00（2.00，14.00） points, respectively.The results of generalized linear regression analysis showed that the main influencing factors for knowledge, attitude, and practice regarding disinfection after intermittent medication administration through NFC were length of clinical service, professional title, department, membership in the venous therapy group, whether being a specialized venous therapy nurse, and relevant training received （all P＜0.05）.Conclusion The knowledge level in nurses on disinfection after intermittent administration through NFC is not high, and their attitude is acceptable but their practice is insufficient.It is necessary to pay attention to the different characteristics of nurses and provide targeted training on disinfection related knowledge and clinical practice after intermittent administration through NFC, striving to prevent the occurrence of catheter-related bloodstream infection.