International Journal of Molecular Medical Science, 2025, Vol.15, No.1, 9-19 http://medscipublisher.com/index.php/ijmms 11 immunosensor for detecting Cyfra 21.1 directly from saliva, demonstrating high sensitivity and specificity in identifying early-stage OSCC (Jafari and Hasanzadeh, 2020). Beyond individual proteins, multiplexed proteomic approaches can enhance the accuracy of early detection. For example, IL-8 has been studied as a biomarker due to its association with inflammation and tumor progression. Xu et al. (2020) reported that combining IL-8 with other salivary proteins in a diagnostic panel significantly improved the ability to detect early OSCC, highlighting the value of protein panels in non-invasive screening methods (Xu et al., 2020). Advanced proteomic technologies, such as high-multiplex aptamer-based platforms, have also been explored for identifying novel serum biomarkers. Blatt et al. (2023) used such a platform to analyze the serum proteome of OSCC patients, identifying several proteins that could serve as early indicators of the disease. This approach underscores the potential of advanced proteomics for creating personalized diagnostic tools that could greatly enhance early detection strategies (Blatt et al., 2023). 2.4 Circulating tumor DNA (ctDNA) Circulating tumor DNA (ctDNA) analysis has gained attention as a promising method for early cancer detection, including OSCC. ctDNA contains genetic mutations, methylation patterns, and other tumor-specific alterations that can be detected in blood samples. Campos-Carrillo et al. (2019) reviewed the utility of ctDNA in early cancer detection, emphasizing its potential for identifying genetic mutations associated with oral cancers and offering a non-invasive alternative to traditional biopsy methods (Campos-Carrillo et al., 2019). However, the low abundance of ctDNA in the early stages of cancer poses a challenge for detection. Phallen et al. (2017) introduced targeted error correction sequencing (TEC-Seq) to improve the sensitivity of ctDNA assays, enabling the detection of low-frequency mutations with high accuracy. This technique has shown promise for identifying early-stage cancer alterations, providing a valuable tool for early screening and monitoring (Phallen et al., 2017). Combining ctDNA analysis with other biomarkers has further enhanced diagnostic performance. Bronkhorst et al. (2019) suggested that integrating ctDNA detection with multi-omic approaches, such as combining genetic and epigenetic markers, could improve sensitivity and specificity, particularly for early-stage tumors. This synergistic approach may offer a more comprehensive view of the tumor landscape, allowing for earlier intervention and better outcomes (Bronkhorst et al., 2019). 3 Technological Advances in Biomarker Detection 3.1 Next-generation sequencing (NGS) Next-Generation Sequencing (NGS) is a revolutionary technology that enables high-throughput analysis of genetic material, allowing for comprehensive profiling of mutations, copy number variations, and other genomic alterations associated with oral cancer. NGS technologies facilitate whole-genome, whole-exome, and targeted sequencing, providing detailed insights into the genetic landscape of Oral Squamous Cell Carcinoma (OSCC). For example, Zhang et al. (2019) discussed the clinical application of NGS in precision medicine, emphasizing its role in identifying cancer driver mutations and enabling personalized treatment strategies for various cancers, including oral cancer (Zhang et al., 2019). NGS has also been applied in the context of liquid biopsy, allowing for the detection of circulating tumor DNA (ctDNA) and mutations in peripheral blood samples. Chen and Zhao (2019) highlighted the use of NGS in liquid biopsy to sequence ctDNA, offering a non-invasive method to monitor tumor dynamics in real time, which is particularly useful for cancers like OSCC where repeated tissue biopsies are challenging (Chen and Zhao, 2019). This method allows for early detection of molecular changes even before clinical symptoms appear, thus facilitating timely interventions. Moreover, targeted NGS panels have been developed specifically for oral cancer research to focus on frequently mutated genes such as TP53 and NOTCH1. Kim et al. (2020) emphasized the role of NGS in identifying novel
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