To establish a convenient, objective and applicable method to assess gingival biotype using periodontal probing.A total of 66 maxillary anterior teeth from 14 volunteers (6 males, 8 females) with healthy gingiva, aged from 20 to 30 years, were recruited in this study. The gingival biotypes were evaluated by whether or not the outline of the periodontal probe was clear inside the gingiva. Gingival thickness of the cemento-enamel junction (CEJ) in upper anterior teeth was measured by cone-beam CT (CBCT).The method of periodontal probing-assessed gingival biotype had good consistency and repeatability. The result of probing-assessed gingival biotype was consistent with that obtained by CBCT. The mean thickness of gingiva in thin, compromised and thick gingival biotype was (1.02 ± 0.20), (1.28 ± 0.25) and (1.46 ± 0.25) mm. These differences were significant for all three comparisons (P = 0.001, respectively).Periodontal probing-assessed gingival biotype is a simple, relatively objective and suitable method for clinical examination.