Study Objective To assess the safety and effectiveness of laparoendoscopic single-site surgery (LESS) compared to conventional laparoscopic surgery for cervicovaginal reconstruction of congenital vaginal and cervical aplasia. Design This was a retrospective study of 19 women who were diagnosed as congenital vaginal and cervical aplasia with a functional endometrial cavity and underwent cervicovaginal reconstruction using small intestinal submucosa graft between January 2013 and December 2018 at the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China. Setting University hospital. Patients or Participants Four women underwent LESS and 15 women underwent conventional laparoscopic surgery for cervicovaginal reconstruction. Interventions In the LESS group, all surgical procedures were performed through a single umbilical multichannel port with 2.5cm incision. In the conventional group, the procedures were completed using a 10-mm umbilical optical trocar and 3 additional 5-mm ancillary trocars. Measurements and Main Results Clinical characteristics, perioperative data, subsequent convalescence and body image were postoperatively assessed. The preoperative characteristics did not significantly differ between the two groups. The median operative time was 212min in the LESS group and 180min in the conventional group (P=0.49). The postoperative hemoglobin drop was similar in the two groups (9.75±7.32g/dl VS 11.67± 8.64 g/dl, P=0.69). There was no complication in the two groups. There was no difference in hospitalization and cost between the two groups. At a median follow-up of 38 (4–64) months, all patients experienced relief of abdominal pain and resumed menstruation. The vaginal-length gain was similar in the two groups (7.25± 0.50cm VS 6.63 ± 0.58 cm, P=0.71). The patients in the LESS group showed higher satisfaction with their body (11.50±1.00 VS 16.07 ± 01.83 cm, P<0.01) and their scar (22.00± 0.82 VS 19.53 ± 2.13cm, P=0.40). Conclusion The single-port technique for cervicovaginal reconstruction is feasible, safe, and equally effective compared to the conventional technique. To assess the safety and effectiveness of laparoendoscopic single-site surgery (LESS) compared to conventional laparoscopic surgery for cervicovaginal reconstruction of congenital vaginal and cervical aplasia. This was a retrospective study of 19 women who were diagnosed as congenital vaginal and cervical aplasia with a functional endometrial cavity and underwent cervicovaginal reconstruction using small intestinal submucosa graft between January 2013 and December 2018 at the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China. University hospital. Four women underwent LESS and 15 women underwent conventional laparoscopic surgery for cervicovaginal reconstruction. In the LESS group, all surgical procedures were performed through a single umbilical multichannel port with 2.5cm incision. In the conventional group, the procedures were completed using a 10-mm umbilical optical trocar and 3 additional 5-mm ancillary trocars. Clinical characteristics, perioperative data, subsequent convalescence and body image were postoperatively assessed. The preoperative characteristics did not significantly differ between the two groups. The median operative time was 212min in the LESS group and 180min in the conventional group (P=0.49). The postoperative hemoglobin drop was similar in the two groups (9.75±7.32g/dl VS 11.67± 8.64 g/dl, P=0.69). There was no complication in the two groups. There was no difference in hospitalization and cost between the two groups. At a median follow-up of 38 (4–64) months, all patients experienced relief of abdominal pain and resumed menstruation. The vaginal-length gain was similar in the two groups (7.25± 0.50cm VS 6.63 ± 0.58 cm, P=0.71). The patients in the LESS group showed higher satisfaction with their body (11.50±1.00 VS 16.07 ± 01.83 cm, P<0.01) and their scar (22.00± 0.82 VS 19.53 ± 2.13cm, P=0.40). The single-port technique for cervicovaginal reconstruction is feasible, safe, and equally effective compared to the conventional technique.