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Background: There is an ongoing research for breast cancer diagnostic tools that are cheaper, more accurate and more convenient than mammography. Methods: In this study, we employed surface-enhanced Raman scattering (SERS) for analysing urine from n = 53 breast cancer patients and n = 22 controls, with the aim of discriminating between the two groups using multivariate data analysis techniques such as principal component analysis&mdash, linear discriminant analysis (PCA-LDA). The SERS spectra were acquired using silver nanoparticles synthesized by reduction with hydroxylamine hydrochloride, which were additionally activated with Ca2+ 10&minus, 4 M. Results: The addition of Ca(NO3)2 10&minus, 4 M promoted the specific adsorption to the metal surface of the anionic purine metabolites such as uric acid, xanthine and hypoxanthine. Moreover, the SERS spectra of urine were acquired without any filtering or processing step for removing protein traces and other contaminants. Using PCA-LDA, the SERS spectra of urine from breast cancer patients were classified with a sensitivity of 81%, a specificity of 95% and an overall accuracy of 88%. Conclusion: The results of this preliminary study contribute to the translation of SERS in the clinical setting and highlight the potential of SERS as a novel screening strategy for breast cancer.