The aim of this case report was to evaluate the results of isthmusectomy and pyeloplasty of horseshoe kidney with the da Vinci robotic-assisted laparoscopy system.This case presented 1 patient with left back pain, associated with lower abdominal pain, and then she underwent the isthmusectomy and dismembered pyeloplasty using robotic-assisted laparoscopy simultaneously. The operation was performed by a transperitoneal approach using 5 ports.We cut the renal isthmus by means of bipolar scissors and then closed the renal parenchyma with 3-0 absorbed stitches. The total operation time was 123 min including simultaneous dismembered pyeloplasty. Blood loss was <50 mL. There were no complications either during or after the procedure. The oral nutrition and mobilization were included on the second day after surgery. The peritoneal drainage was removed on the eighth day. Long-term follow-up after treatment showed good results.The da Vinci robotic-assisted laparoscopy is an alternative to open surgery and laparoscopy, particularly in the correction of congenital defects of the urinary tract. Furthermore, the da Vinci robotic-assisted laparoscopy technique in isthmusectomy and pyeloplasty is safe for patient as shown by our results.