High-Accuracy Renal Cell Carcinoma Discrimination through Label-Free SERS of Blood Serum and Multivariate Analysis

Renal cell carcinoma (RCC) represents the sixth most frequently diagnosed cancer in men and is asymptomatic, being detected mostly incidentally. The apparition of symptoms correlates with advanced disease, aggressive histology, and poor outcomes. The development of the Surface-Enhanced Raman Scattering (SERS) technique opened the way for investigating and detecting small molecules, especially in biological liquids such as serum or blood plasma, urine, saliva, and tears, and was proposed as a simple technique for the diagnosis of various diseases, including cancer. In this study, we investigated the use of serum label-free SERS combined with two multivariate analysis tests: Principal Component Analysis combined with Linear Discriminate Analysis (PCA-LDA) and Supported Vector Machine (SVM) for the discrimination of 50 RCC cancer patients from 45 apparently healthy donors. In the case of LDA-PCA, we obtained a discrimination accuracy of 100% using 12 principal components and a quadratic discrimination function. The accuracy of discrimination between RCC stages was 88%. In the case of the SVM approach, we obtained a training accuracy of 100%, a validation accuracy of 92% for the discrimination between RCC and controls, and an accuracy of 81% for the discrimination between stages. We also performed standard statistical tests aimed at improving the assignment of the SERS vibration bands, which, according to our data, are mainly due to purinic metabolites (uric acid and hypoxanthine). Moreover, our results using these assignments and Student's t-test suggest that the main differences in the SERS spectra of RCC patients are due to an increase in the uric acid concentration (a conclusion in agreement with recent literature), while the hypoxanthine concentration is not statistically different between the two groups. Our results demonstrate that label-free SERS combined with chemometrics holds great promise for non-invasive and early detection of RCC. However, more studies are needed to validate this approach, especially when combined with other urological diseases.