A Rapid Detection of Nipah Virus Nucleocapsid Antigen in Inactivated Viral Cultures

作者：최경무, Jung Kim, Hansaem Lee, Pyeonghwa Jeon, Jae Hun Jeong, Yujin Jung, Jin Seo Park, Mi Kyoung Lee, Kyung-Chang Kim, Joo-Yeon Lee, Ha Yeon Park, Changwoo Park, Seil Kim, Hana Yi, Jihwan Song, Hyun-Joo Kim, Jong-Hwan Lee, Hong Gi Kim · 发表于：Figshare · 年份：2026 · DOI：10.6084/m9.figshare.33183998.v1 · 研究领域：Virology and Viral Diseases、Respiratory viral infections research、Biosensors and Analytical Detection

Nipah virus (NiV) is a highly lethal zoonotic henipavirus that causes severe respiratory disease and encephalitis and remains a priority pathogen for outbreak preparedness. Timely diagnosis is essential for containment, yet nucleic acid–based testing is often impractical in outbreak-prone, resource-limited settings. Here, we report the development of a rapid antigen test for NiV based on a cellulose nanobead (CNB)–enhanced lateral flow immunoassay (LFIA) targeting the viral nucleocapsid protein (NP). Anti-NiV NP antibodies were isolated from human synthetic phage-display libraries (scFv and Fab) by iterative bio-panning, reformatted as full-length IgGs, and screened using ELISA and immunofluorescence assays. Four lead antibodies were advanced to a sandwich-pairing matrix, which identified an optimized capture–detection configuration (mAb #5 as capture; mAb #2 as detection). Bio-layer interferometry confirmed nanomolar binding affinities for the selected antibodies (K D = 39.50 nM for mAb #5; K D = 26.36 nM for mAb #2). The finalized CNB-LFIA produced results within 15 min and achieved an analytical limit of detection of 1 ng/mL for recombinant NiV NP. The assay observed no cross-reactivity with SARS-CoV-2, SARS-CoV, MERS-CoV, or influenza A, whereas weak cross-reactivity for measles virus NP was observed only at a high antigen concentration (2 µg/mL). Using γ-irradiated cultured NiV quantified by RT-ddPCR, the LFIA detected viral load down to 4.1 × 10 7 copies/mL (4.1 × 10...