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Background: Recently, cytology has been applied to the diagnosis of oral lesions. We aimed to explore the diagnostic accuracy of oral cytology based on the histological diagnosis. Methods: Histological diagnoses of 327 cases were classified as Negative, Borderline lesion -, Borderline lesion +, oral intraepithelial neoplasia/carcinoma in situ (OIN/CIS), or Positive. Cytological diagnoses were classified as NILM (negative for intraepithelial lesion or malignancy), LSIL (low-grade squamous intraepithelial lesion), HSIL (high-grade squamous intraepithelial lesion), or SCC (squamous cell carcinoma). The cytology slides were evaluated by 10 raters and the results were compared with the histology results. Results: In 142 cases that were histologically negative, the number of NILM, LSIL, HSIL, and SCC and other malignancy was 77 (54.2%), 47 (34.3%), 8 (5.6%), and 10 (7.0%), respectively. Among 32 cases of Borderline lesion -, the number of NILM, LSIL, HSIL, and SCC and other malignancy was 11 (34.3%), 11 (34.3%), 9 (28.1%), and 1 (3.1%), respectively. Also, in 4 cases of Borderline lesion +, the number of NILM, LSIL, HSIL, and SCC and other malignancy was 2 (50.0%), 0 (0.0%), 0 (0.0%), 2 (50.0%), respectively. Among 12 cases of OIN/CIS, the number of NILM, LSIL, HSIL, and SCC and other malignancy was 1 (8.3%), 2 (16.7%), 4 (33.3%), and 5 cases (41.7%), respectively. Among 137 cases with a histological diagnosis of Positive, the number of NILM, LSIL, HSIL, and SCC and other malignancy was 7 (5.1%), 22 (16.1%), 19 (13.9%), and 89 (65.0%), respectively. Sensitivity, specificity, and positive predictive and negative predictive values were 93.5, 50.6, 62.4, and 89.8%, respectively, when the cytological diagnosis of Negative was assumed to be NILM; they were 77.8, 83.9, 81.0 and 81.1%, respectively, if the cytological diagnosis of Negative was assumed to be NILM and LSIL. The number of false-positive and false-negative diagnosis affected cases with LSIL and HSIL may indicate the difficulty in the cytological diagnosis of borderline lesions. While the negative predictive value was relatively high (89.8%) when cytological Negative was assumed to be NILM only. Conclusion: Histopathological examination should be recommended in cases with cytological diagnoses of LSIL, HSIL, and SCC. [ABSTRACT FROM AUTHOR]