Objective To investigate the level of and factors associated with illness perception in patients after limb amputation, and to provide reference for targeted intervention. Methods A cross-sectional survey was conducted in 188 patients after limb amputation by employing a general data questionnaire, the Barthel Index, Illness Perception Questionnaire, and the Hospital Anxiety and Depression Scale. Results The samples scored 50.57±9.11 on illness perception, 65.08±13.30 on the Barthel index, 14.38±3.70 on anxiety and 15.12±3.20 on depression. Multiple linear regression analysis showed that amputation sites, anxiety and depression, hospital stay length and self-care ability were the main influencing factors of illness perception (adjusted R square=0.442,P＜0.05,P＜0.01). Conclusion Illness perception in patients after limb amputation is at moderate to above average level, and is affected by surgical site related functional changes and mental state of the patient. Clinical nurses should pay attention to and provide targeted interventions to improve their recovery and illness perception.