Cancer is the disease found to be the reason for the largest portion of deaths in the world annually and these mortality values are expected to increase in the future. Early detection of cancer biomarkers may help save millions of lives, particularly by implementing non-invasive and economical detection methods. In this review, we tabulated and quantitatively compared the data collected in 173 rows from 124 publications, which describe the clinical application of various methods in detection of cancer biomarkers. Those methods include mass spectrometry (MS), immunoassays (IAs), enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), surface-enhanced Raman spectroscopy (SERS), and Fourier-transform infrared spectroscopy (FTIR). We found that direct methods may have an advantage over indirect methods. Direct SERS reported in clinical applications can also achieve a higher area under the curve, higher sensitivity, and specificity than those parameters for ELISA, PCR, MS, and FTIR applications. Based on the average area under the curve (AUC) values reported in the last 6–7 years for each method, the performance of the analytical methods for the clinical cancer detection increases from IAs (0.76), ELISA (0.83), MS (0.87), and PCR (0.89) to FTIR (0.95) and SERS (0.97).

Human saliva is a heterogeneous bodily fluid with a complex composition, which contains antibodies, proteins, and viruses, making it applicable in clinical diagnosis. There are several advantages of the analysis of saliva samples over other biofluids, including a non-invasive and simple collection procedure for extraoral detection. Biomarker or pathogen detection in saliva can be performed with various methods: mass spectrometry, PCR, ELISA, electrochemical, and optical methods such as fluorescence, SPR, and SERS. The early detection of cancer and other disease biomarkers, as well as infectious agents, can be crucial for effective treatment and minimization of mortality from those diseases. The following paper reviews extraoral detection techniques to identify the most sensitive methods for diagnosing early and asymptomatic patients. The LODs collected and tabulated from 149 analytical papers, alongside the sensitivity, specificity, and sometimes the area under the curve (AUC) tabulated from 118 clinical studies, have all become parameters for the comparative quantitative analysis. Based on the limited but substantial number of analytical studies on the detection of cortisol in saliva (29), the electrochemical platforms demonstrated the highest sensitivity, with a geometric mean LOD of 11 pM. Within these methods, voltametric ones showed the best performance with 6 pM geometric mean LOD. Electrochemical techniques are then followed by immunoassay- and mass spectrometry-based platforms, with corresponding geometric average LOD values of 39.1 and 171 pM, respectively. However, clinical outcomes are at least as meaningful as LOD values. In terms of clinical analysis, ELISA and direct-SERS outperformed other methods, achieving balanced accuracy of approximately 87% and AUC values of 0.96 for direct SERS and 0.86 for ELISA. MS and PCR followed closely, with balanced accuracies around 84%. While the direct SERS is not yet widespread in clinical applications, its potential can be forged if the standardization issue is addressed.