Abstract: Objective  To investigate the current status of standardized operation of maintenance hemodialysis by autologous arteriovenous fistula puncture in hemodialysis nurses.  Methods  In July to August 2023, the videos of hemodialysis operation by autologous arteriovenous fistula puncture were collected nationwide through the platform of “Comprehensive Dialysis Management Capacity Building Project, National Health Commission”. Two experts reviewed the videos by using a self-made scoring scale that contains  five dimensions: instrument and occupational protection, hand hygiene timing and method, pre-operation assessment and inspection, vascular puncture and drug injection, and safety verification, and the reasons for point deduction were recorded. Differences of scores were compared among hospital levels, whether or not from a blood purification teaching base, and by different professional titles.  Results  A total of 419 valid videos were collected. The scoring rates of the five dimensions were as follows: instrument and occupational protection (85.232%), hand hygiene timing and method (84.417%), pre-operation assessment and inspection (74.213%), vascular puncture and drug injection (77.513%), and safety verification (68.702%). Total scores were higher from tertiary hospitals and above than from other hospitals (t=3.466, P=0.015); total scores were higher from blood purification teaching bases than from non-teaching bases (t=5.062, P=0.004); total scores had no significant difference from nurses of different professional titles (F=0.292, P=0.747).  Conclusion  The scoring rates of the five dimensions ordered from low to high are as follows: safety verification, pre-operation assessment and inspection, vascular puncture and drug injection, hand hygiene timing and method, and instrument and occupational protection, suggesting the key points of hemodialysis operation training in the future. The operation quality is relatively lower in the hospitals below the tertiary level and non-teaching bases, highlighting the hemodialysis units we should concern with and pay assistance to.