Renal Clearable Luminogenic Reporter for Ultrasensitive Influenza Virus Imaging and Efficient Antiviral Therapies Monitoring in Living Mice

ABSTRACT Influenza, which causes respiratory tract infections and related complications, poses a major threat to global public health. However, the rapid and accurate detection of influenza viruses in controlling the flu pandemic remains challenging, as current diagnostic methods are static and unable to distinguish between viable and nonviable virus or directly monitor viral replication dynamics. Herein, we report viral luminogenic reporters (VLRs) with chemiluminescence/fluorescence dual‐response for non‐invasive imaging and urinalysis of the H1N1 virus protease. VLR comprises a bicyclic dioxetane chemiluminophore signaling scaffold caged by a N‐acetylneuraminic acid, which further hooks a renal clearable moiety (2‐hydroxypropyl)‐β‐cyclodextrin. VLR achieves a limit of detection of 2.62 CCID ₅₀ /mL in H1N1 virus detection, which was 10.4‐fold and 529.8‐fold lower than that of ICA‐qPCR and PMA‐qPCR assays. After intratracheal administration into H1N1 virus‐infected mice, VLR can efficiently accumulate in the lungs and specifically react with neuraminidase to restore its near‐infrared chemiluminescent/fluorescent signals for real‐time imaging. Leveraging the renal clearance (∼94% ID), VLR allows for remote detection of H1N1 virus infections and monitoring of antiviral therapeutic efficacy through in vitro urinalysis. Therefore, this study highlights a significant advance in addressing the critical gap in dynamic monitoring of virus replication activity and transforming virus‐specific probes into urinary reporters.