Aim. The aim of this study is to evaluate the diagnostic value of machine learning- (ML-) based quantitative texture analysis in the differentiation of benign and malignant thyroid nodules. Materials and methods. A sum of 306 quantitative textural features of 235 thyroid nodules (102 malignant, 43.4%; 133 benign, 56.4%) of a total of 198 patients were investigated using the random forest ML classifier. Feature selection and dimension reduction were conducted using reproducibility testing and a wrapper method. The diagnostic accuracy, sensitivity, specificity, and area under curve (AUC) of the proposed method were compared with the histopathological or cytopathological findings as reference methods. Results. Of the 306 initial texture features, 284 (92.2%) showed good reproducibility (intraclass correlation ≥0.80). The random forest classifier accurately identified 87 out of 102 malignant thyroid nodules and 117 out of 133 benign thyroid nodules, which is a diagnostic sensitivity of 85.2%, specificity of 87.9%, and accuracy of 86.8%. The AUC of the model was 0.92. Conclusions. Quantitative textural analysis of thyroid nodules using ML classification can accurately discriminate benign and malignant thyroid nodules. Our findings should be validated by multicenter prospective studies using completely independent external data.