Objective  This study aimed to analyze and explore the factors influencing the presence of late thrombosis in autogenous arteriovenous fistula (AVF) in patients on maintenance hemodialysis (MHD).  Methods  In this cross-sectional study, Clinical data of the patients who received MHD at the Hemodialysis Vascular Access Clinic, The Central Hospital Affiliated to Shandong First Medical University between January 2023 and December 2023 were collected. Patients were divided into a patency group and an obstruction group based on the presence or absence of thrombosis in AVF. Binary logistic regression was used to explore the potential influencing factors.  Results  A total of 521 MHD patients were included in this study, among whom thrombosis in AVF occurred in 252 cases (48.37%). Binary logistic regression analysis showed that compression time for hemostasis (OR=8.145, 95% CI: 3.543～18.727, P＜0.001), fistula functions (including insufficient blood flow during hemodialysis, high venous pressure, and puncture failure) in the latest month (OR=47.404, 95% CI: 11.127～201.949, P＜0.001), pre-dialysis systolic blood pressure (OR=0.971, 95% CI: 0.949～0.993, P=0.011), pre-dialysis diastolic blood pressure (OR=1.111, 95% CI:1.076～1.148, P＜0.001), minimum diastolic blood pressure during hemodialysis (OR= 0.923, 95% CI: 0.886～0.961, P＜0.001), regional cannulation (OR=66.674, 95% CI: 7.505～592.350, P＜0.001), buttonhole cannulation (OR=18.578, 95% CI: 6.154～56.086, P＜0.001), and a history of AVF dysfunction for two or more two times (OR=3.967, 95% CI: 1.614～9.751, P=0.003) were the risk factors influencing the presence of thrombosis in AVF.  Conclusion Compression time for hemostasis ≥30 minutes, AVF dysfunction in the latest month, higher pre-dialysis diastolic blood pressure, and a history of multiple AVF failures were the risk factors for late thrombosis in AVF, while rope ladder cannulation, high pre-dialysis systolic blood pressure, and high minimum diastolic blood pressure during hemodialysis were the protective factors against late thrombosis in AVF.