Objective To evaluate the safety and efficacy of ultrasound-guided one supracostal upper-calyx percutaneous access nephrolithotomy for staghorn stone. Methods A total of 581 patients with staghorn stone were treated with one supracostal upper-calyx access percutaneous nephrolithotomy from October 2004 to October 2011. The maximal diameters of stone measured by plain abdominal radiography were 3.6-6.5 cm, with a mean of (5.10±0.82) cm. Target calyx was posterior upper-calyx,puncture point was selected in tenth or eleventh intercostal space as guided by ultrasound, and the F22-24 upper-calyx percutaneous access was established. 17F nephroscope and 70 W holmium laser were employed to crush the stones. The operation time, perioperative complications, and stone-free rate were recorded. Results The upper-calyx percutaneous access was established successfully in all patients. The mean operation time was (82.07±16.25) min (range 54 to 140 min). The stone-free rate was 85.4% (496/581) after first-stage procedure. Fourteen patients (2.4%) had postoperative high fever, and 3 of them (0.5%) had infectious shock. Four patients received blood transfusion, and 2 of them underwent digital subtraction angiography to control bleeding. Four patients (0.7%) underwent thoracic drainage. There were no injuries to the lung, spleen, colon, or liver. The overall postoperative complication rate was 3.8%. The mean postoperative hospital stay was (5.55±1.35) days (ranging 4-15 days). Conclusion One supracostal upper-calyx percutaneous access can achieve high stone-free rate in treating staghorn stone, but with relatively high risk of complications. Ultrasound-guided technique can greatly reduce complication, making the procedure safer.