This study aims to evaluate three types of manual toothbrushes [Cross action (CA), Flat trim (FT), and orthodontic type (OT)] in terms of efficacy in plaque removal in patients undergoing fixed orthodontic treatment. Manual toothbrushes are an essential part of oral hygiene for primary prevention. Plaque control, however, can be influenced by a number of individual and material-related factors. Individual factors include the presence of fixed orthodontic appliances on tooth surfaces, such as brackets and bands, which create difficulties with oral hygiene and lead to plaque formation. The evidence for the effectiveness of advanced bristle designs (multilevel, criss-cross) of the manual toothbrush alone in removing plaque in patients undergoing orthodontic therapy is limited. The experiment followed the Consolidated Standards of Reporting Trials (CONSORT) guidelines. This was a three treatment, three-period crossover clinical trial with a single brushing exercise. Thirty subjects were randomized to one of the three treatment sequences of different bristle designs: (CA, FT, and OT). The primary outcome measure was the difference in the plaque scores (baseline minus post-brushing) at each study period, as determined by the Turesky-Modified Quigley-Hein Plaque Index. Of the thirty-four subjects enrolled in the study, thirty of the subjects met the inclusion criteria and completed all three periods of the study. The mean age was 19.5 ± 1.52 years, with a range of 18-23 years. The differences between treatments in plaque score reduction after brushing were statistically significant (p-value < .001). The treatment differences were statistically significant (p-value < .001) favoring the FT toothbrush over the OT and CA types of toothbrush designs. On the contrary, the difference between the OT and CA types was not statistically significant. Plaque was significantly removed by the conventional FT toothbrush after a single brushing compared to the OT and CA types.