Background. Pain due to vascular needle insertion has been reported in 40–60% of hemodialysis (HD) patients. Evidence suggests that there is typically no single method for relieving the pain of inserting vascular needles in HD patients. This study aimed to compare the effectiveness of EMLA cream and Valsalva maneuver (VM) on pain severity during vascular needle insertion in HD patients. Methods. This randomized, controlled, clinical trial was conducted on 90 patients undergoing hemodialysis in the hemodialysis unit of Kowsar Hospital, affiliated with Semnan University of Medical Sciences, in Semnan, Iran. Patients were selected via convenience sampling and were randomly assigned to one of the three groups (EMLA, VM, and control groups). For the patients in the EMLA group, 2.5 g of EMLA cream was applied 60 minutes before the start of dialysis. For patients in the VM group, a maneuver was performed for 16–20 seconds before the needle was inserted. Patients in the control group received only routine care without any additional interventions. The pain severity in the three groups was measured using the visual analog scale (VAS) two minutes after vascular needle insertion. Results. The results showed that the mean pain severity during cannulation was 2.06 ± 2.19 in the EMLA group, 3.2 ± 30.42 in the VM group, and 6.20 ± 1.49 in the control group, suggesting a significant difference between the groups (
P
<
0.001
). Pairwise comparison of the mean pain severity showed that it differed significantly in the EMLA and VM groups from the control group (
P
<
0.001
), but no significant difference was found between the EMLA and VM groups (
P
=
0.067
). Conclusion. According to the results, EMLA cream was as effective as VM in reducing the pain severity caused by arteriovenous fistula (AVF) cannulation. Therefore, the use of EMLA cream and VM is recommended for reducing the severity of AVF cannulation pain. Trial Registration. Iranian Registry of Clinical Trials, Trial No : IRCT20120109008665N12, registered on 3 June 2020.