To explore an in vitro suture-training mode in laparoendoscopic single-site surgery (LESS) with training box, different access ports and suture modes based on clinical experiences.

The suture procedure was performed by one experienced surgeon using one-handed and two-handed methods. The suture included a fixed and a mobile target. The time of 30 sutures in 4 ports was compared using the two suture methods and two targets. The control group was suture without port. The followed suture practice was finished by 5 other surgeons without LESS experience once a week without port.

The suture training was successfully completed with a modified conventional laparoscopic training box. When suturing the fixed target, the one-handed method was less time-consuming than the two-handed one (P<0.05). When suturing the mobile target, the two-handed method was less time-consuming than the one-handed one. Suture with a bigger port was more time-consuming. There was no difference in suture time with or without a port. The suture time was gradually shortened in the first 3 times [(16.5±0.2) s, (13.4±0.3) s, (12.0±0.1) s](P<0.05) and there was no statistical difference between the 4th, 5th week and the 3rd week[(12.3±0.2) s, (12.3±0.2) s](P>0.05).

The suture training in LESS could be conducted with a modified conventional laparoscopic training box without port. One-handed suture should be emphasized and a large port is not suitable for training. Practices once a week for 3 weeks would be appropriate for the training.