Development and Clinical Evaluation of a Nanoparticle-based Biosensor for Rapid Detection of Hepatitis E Virus in Human Serum

Hepatitis E virus (HEV) genotype 3 is a zoonotic pathogen increasingly recognized in industrialized countries that is often linked to foodborne or animal contact transmission. While most infections are self-limited, outbreaks pose a significant public health challenge, especially in immunocompromised populations and blood donation recipients. Early-stage viral detection is critical for effective disease management and containment, since it helps to identify clusters of infection and implement public health measures to control the spread of the disease, especially in community settings, which underscores the need for innovative rapid through-put diagnostic platforms. In this study, we present the development and comprehensive analytical/clinical validation of a novel nanoparticle-based genetic biosensor, enabling rapid HEV detection directly from human serum with a turnaround time of approximately five minutes. The system consists of three individual kits, each containing nanoparticles functionalized with validated nucleic acid probes targeting conserved HEV genomic regions. When tested on clinical samples positive for HEV-3, the biosensor showed full concordance with RT-qPCR, detecting viral loads as low as <1 IU/mL. Moreover, it is a highly specific system when tested against other known human virus, namely hepatitis A, hepatitis B, cytomegalovirus and, Epstein-Barr virus. The platform is portable, cost-effective, and does not require nucleic acid extraction or amplification steps, making it highly suitable for field settings or Point-of-Care Testing (PoCT). This rapid and accessible approach holds promise for expanding diagnostic capabilities, particularly in resource-limited or field settings, where it can have a great impact on early outbreak detection, rapid diagnosis of infections, tracking disease spread, and assessing food and water safety. The development of rapid, in field tests are crucial for enabling prompt public health responses, and thus, support the global effort to eliminate viral hepatitis.