LAMP-based electrochemical platform for monitoring HPV16 genome integration.

Human papillomaviruses (HPVs) represent a diverse family of double-stranded DNA viruses implicated in epithelial tissue infections and various cancers, notably cervical cancer [1]. Integration of high-risk HPV types, such as HPV16 and HPV18, into the host genome, primarily near common fragile sites, contributes to genomic instability and cancer progression [2]. Traditional methods for detection of HPV integration, like immunohistochemistry and PCR-based assays, are hindered by technical challenges and complex workflows [3]. Newly emerging isothermal amplification techniques (IATs) combined with electrochemical (EC) detection platforms offer promising alternatives for rapid and cost-effective HPV analysis [4]. Here, we introduce a novel biosensing platform that combines loop-mediated amplification (LAMP) with EC analysis specifically targeting HPV16 integration, the most prevalent oncogenic genotype. By targeting oncoviral mRNAs E2 and E7, our method facilitates HPV integration analysis. Utilizing cell lines with known HPV16 integration status, EC readout revealed distinct E7 expression patterns in HPV16-positive and negative cell lines. Furthermore, E2 mRNA expression correlated with integration status, validating the biosensor's reliability. Finally, we evaluated HPV16-positive cervical tissue samples to assess viral integration status, obtaining clear distinct E2 and E7 mRNA signals between episomal and integrated virus forms, showcasing its applicability and robustness in clinical samples. This innovative strategy holds promise for advancing HPV diagnostics, offering valuable information whether women with precancerous lesions are at higher risk of disease progression.