Objective To explore the application of multiple needle puncture technique in maintenance hemodialysis (MHD). The clinical value of autologous arteriovenous fistula (AVF) puncture in patients with MHD, analyze the impact of this technique on vascular complications, puncture pain, and nursing effectiveness, and optimize the long-term standardized puncture management of AVF. Methods Patients with AVF who were enrolled in the Department of Nephrology of the First People’s Hospital of Longquanyi District Chengdu in November 2024 were selected. Patients were categorized into the multiple needle puncture group and the rope ladder puncture group according to puncture methods. All enrolled patients completed the 18-month follow-up period, with a follow-up deadline of May 2026. The incidence of AVF vascular complications (vascular stenosis, aneurysm, thrombus), puncture pain scores, and the one-time puncture success rate of junior nurses were analyzed between the two groups at 1, 6, 12 and 18 months after intervention. ResultsA total of 102 patients were included, with 51 in each group. The results of generalized estimating equations showed that there were time effects, inter-group effects, and time-group interaction effects on vascular access complications of fistula between the two groups (P<0.05). Compared with the rope ladder puncture group, the multiple needle puncture group exhibited a lower incidence of fistula vascular stenosis, aneurysm and thrombus, and puncture pain score (P<0.05). In addition, the one-time puncture success rate of junior nurses was higher (P<0.05). Conclusions Multiple needle puncture can protect the blood vessels of internal fistula, alleviate puncture-related pain, reduce the risk of vascular access-related complications, maintain the long-term smoothness of internal fista, and improve the effectiveness of clinical care. However, this technique has some shortcomings, so the clinical choice of a puncture regimen needs to be tailored to the patient’s vascular conditions.