Abstract: Regional Test is the basis for production test and new variety registration. To overcome the limitation of traditional cultivar evaluation and enhance the competitiveness of new cultivar Gossypium barbadense L. on screening, promotion, demonstration and utilization can provide a scientific basis for setting variety breeding goals and mining high yield potential. Identical and different analysis method was used for comprehensive evaluation of 14 traits in 27 Gossypium barbadense L. cultivars (lines), and cultivars with good comprehensive trait were selected.‘H3549’had the highest comprehensive identical degree, which indicated that‘H3549’ had the best comprehensive traits, 96.79% of its 14 traits reached the breeding goals. The second was‘33687’. The yield and quality traits of the above two cultivars had relative advantage, the comprehensive traits were better, so the two could be preferentially popularized cultivars. Elongation rate, tenacity, yield and boll number per plant of‘K-379’,‘ X-2038’,‘H5161’,‘TH-314’,‘DJ10149’,‘G-92’,‘JSH12-3’,‘Yuanlong-10’ and‘Tarim09- 164’were lower, micronaire value were higher, and improvement approaches had been proposed. Connection degree of‘DJ09-172’,‘Changfeng-2’,‘Tarim08-362’,‘Tarim09-324’,‘Tarim10- 280’and‘11-616’were low with ideal cultivars, and need further observation or to be eliminated. Identical and different analysis method is simple in calculation and easy to master, feasible in regional test of Gossypium barbadense L..