Objective To systematically evaluate the efficacy and safety of Single-incision thoracoscopic surgery (SITS) and two-port video-assisted thoracoscopic surgery (2P-VATS) in the treatment of spontaneous pneumothorax. Methods The databases of CNKI, PubMed, The Cochrane Libray, Web of Science, EMbase, Wanfang and Chinese Medical Association were searched by computer. Literature on SITS treatment of spontaneous pneumothorax from the establishment of the database to March 2023. The data are processed with RevMan 5.4.1. Results Finally, 107 studies were included, including 35 RCTs, 2 cohort studies, and 70 case-control studies. Meta analysis results show that compared to 2P-VATS and three port video assisted thoracoscopic surgery (3P-VATS), SITS had a shorter surgical time [SMD=–0.53, 95%CI (–0.90, –0.16), P=0.005], less intraoperative bleeding [SMD=–1.58, 95%CI (–1.93, –1.22), P<0.000 01; SMD=–1.59, 95%CI (–2.03,–1.14), P<0.000 01], shorter postoperative hospitalization time [SMD=–1.05, 95%CI (–1.29,–0.82), P<0.000 01; SMD=–1.08, 95%CI (–1.39,–0.77), P<0.000 01], and shorter postoperative drainage (catheterization) time [SMD=–0.75, 95%CI (–1.00,–0.50), P<0.000 01; SMD=–1.23, 95%CI (–1.72,–0.75), P<0.000 01], fewer postoperative complications [OR=0.34, 95%CI (0.26,0.45), P<0.000 01; OR=0.47, 95%CI (0.33,0.68), P<0.000 1], fewer postoperative recurrences [OR=0.50, 95%CI (0.33,0.75), P=0.000 8], and lighter postoperative pain [SMD=–1.71, 95%CI (–1.98,–1.45), P<0.000 01; SMD=–2.02, 95%CI (–2.46,–1.59), P<0.000 01]. Compared with 3P-VATS, 2P-VATS had less intraoperative bleeding [SMD=–1.02, 95%CI (–1.81,–0.22), P=0.01] , shorter postoperative hospitalization time [SMD=–0.59, 95%CI (–1.11,–0.06), P=0.03], shorter postoperative drainage (catheterization) time [SMD=–0.46, 95%CI (–0.85,–0.08), P=0.02], fewer postoperative complications [OR=0.36, 95%CI (0.22,0.59), P<0.000 1] , and lighter postoperative pain [SMD=–0.80, 95%CI (–1.08,–0.53), P<0.000 01]. Conclusion SITS and 2P-VATS are an effective and safe method for the treatment of spontaneous pneumothorax and worthy of further promotion and application in clinical practice. Due to limitations in the quantity and quality of included studies, the above conclusions require more large-sample, high-quality studies to be verified.