Background:The life expectancy and prognosis of patients suffering from oral cancer will be considerably improved if oral cancer is detected and treated early. To augment clinical examination and improve diagnosis, adjunctive diagnostic aids such as vital staining have been introduced. Aim:The present research examined the diagnostic efficacy of acetic acid (2%), lugol's iodine (3% dilution), and toluidine blue (1%), to diagnose oral leukoplakia. Materials and Methods:A cross-sectional study was undertaken at a hospital with 45 randomly selected subjects who had clinically verified cases of oral leukoplakia. Acetic acid, lugol's iodine, and toluidine blue were applied in respective order on oral lesions. The results of each staining were then compared to the clinical and histopathologic diagnoses. A control group was made up of cases diagnosed with epithelial hyperplasia. Numbers and percentages are used to present the information. To compare the effects of toluidine blue application, toluidine blue, and acetic acid, the Chi square test was utilised. In terms of its utility in predicting the dysplastic character of the lesion, the sensitivity and specificity test was used to determine diagnostic efficiency and reliability. Results:Toluidine blue staining and lugol's iodine staining both had a sensitivity of 91.59 percent, while the former had a specificity of 23.33 percent and the latter had a specificity of 12.22 percent. The sensitivity of the acetic acid test was 58.25 percent and the specificity was 34.44 percent. There was no statistically significant difference between the staining with three agents and the histopathologic variations in multiple comparisons.