Objective To analyze the sperm DNA fragmentation in different degrees of varicocele (VC) infertility patients after high ligation of the spermatic vein. Methods There were 57 patients with different degrees of VC infertility patients. 27 patients had been diagnosed with level two VC, and 30 patients had been diagnosed with level three VC. All patients' sperm DNA fragmentation index (DFI) were detected and calculated at preoperative one month and postoperative three months. Results 57 VC infertility patients were all suffered from laparoscopic high ligation of the spermatic vein on both sides. There was no statistically difference on preoperative sperm DFI between level two and level three patients (P > 0.05). All patients' sperm DFI were decreased after laparoscopic (P < 0.05), and level two VC infertility patients had significant greater progress than level three patients (P < 0.05). Conclusion Laparoscopic high ligation of the spermatic vein can effectively improve VC infertility patients sperm DFI. It would be better for level two VC infertility patients.