Biofunctional Molecular Large Surfaces Enabling Ultrasensitive Detection for Early Diagnosis of Cervical Cancer

Abstract Human Papillomavirus (HPV) infections are among the leading causes of cervical cancers globally. The World Health Organization has established global targets aimed at reducing the spread of HPV and its associated disease burden. To date, the detection of HPV infections relies primarily on genomic assays targeting viral DNA or mRNA, which, although highly sensitive, require time-consuming PCR protocols performed by trained personnel. In addition, these approaches may be challenged in biological matrices, such as saliva, where viral concentrations are low (e.g., head-neck HPV cancer) or early stage infections. Therefore, alternative strategies enabling rapid, sensitive, and direct detection of viral biomarkers are highly desirable. In this study, we present the Single-Molecule-with-a-Large-Surface (SiMoLS) technology for the detection of the HPV-16 L1 capsid protein in both model and clinically relevant samples added with HPV-16 L1. The SiMoLS platform exploits a biofunctionalized gate electrode integrated within an electrolyte-gated organic field effect transistor, enabling label-free detection at the single- or few-molecules level. By targeting HPV-16 L1, a structural protein abundantly expressed during productive infection, the platform provides direct evidence of viral presence without the need for nucleic acid amplification. The device demonstrates high sensitivity and selectivity across a broad concentration range, extending down to 10–20 M with a confidence level of 99%, while PCA shows a well-defined clustering pattern that separates positive from negative swab real samples in an unsupervised manner. The SiMoLS platform (i) offers rapid analysis with ultrahigh sensitivity, (ii) it is compatible with low sample volumes, supporting its application in point-of-care (POC) settings due to its scalability and ease of use, and (iii) represents a promising tool for the early and decentralized HPV highly sensitive yet reliable screening, particularly in scenarios where conventional methods face limitations.