Purpose: To test the plaque-removal efficacy of a single-tufted toothbrush on the posterior molars compared with a flat-trimmed toothbrush. Methods: Forty-nine subjects were selected. Professional instruction and written brushing instructions were given. After thorough supra-gingival scaling and polishing, all subjects were asked to abstain from oral hygiene procedures for 24 hours prior to the first experiment. The subjects were randomized to a treatment sequence. The modified Quigley and Hein plaque index was recorded pre- and post-tooth brushing, at 6 surfaces of the posterior molars. After a wash-out period, all the remaining plaque was removed professionally. Twenty-four hours of brushing abstinence was again performed. The plaque index was recorded pre- and post-tooth brushing after the subjects were given the second toothbrush in the cross-over sequence. Results: The percentage reductions in plaque scores achieved with the single-tufted brushes were significantly higher than those of the flat-trimmed brush at the maxillary buccal interproximal, marginal and mandibular lingual interproximal site. The other locations showed no significant difference. Conclusions: The results of the present study implied that the single-tufted brush could be an effective tool for the removal of plaque at some, but not all, sites of the posterior molars.