Conserved nucleocapsid epitopes of Influenza A and B viruses in Asia for broad spectrum diagnostics | ||||
| Microbes and Infectious Diseases | ||||
| Articles in Press, Accepted Manuscript, Available Online from 19 July 2025 | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/mid.2025.397204.2941 | ||||
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| Authors | ||||
Radhakrishna muttineni  1; Siripuram Jhansi Rani2; Veena Madhuri K1; Rajitha Jannapureddy3
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| 1Virus Research Laboratory, Department of Zoology, Osmania University, Hyderabad, India, | ||||
| 2Department of Zoology, Veranari Chakali Ilamma Women University, Hyderabad, India | ||||
| 3Retroindica Diagnostics and Therapeutics, Hyderabad, India | ||||
| Abstract | ||||
| Background: Influenza A and B viruses are major drivers of global respiratory morbidity and mortality, highlighting the urgent need for rapid, subtype-independent diagnostics. Conventional assays often target hemagglutinin (HA) or neuraminidase (NA), which undergo frequent antigenic drift, compromising sensitivity during seasonal and pandemic shifts. Objectives: To identify highly conserved, immunogenic, and diagnostically relevant B-cell epitopes within the nucleoprotein (NP) of circulating Influenza A and B strains across Asia and evaluate their suitability for multiplex lateral flow assays (LFAs). Methods: We retrieved 4,592 Influenza A NP sequences and 784 Influenza B NP sequences from NCBI and GISAID. Multiple sequence alignment using MUSCLE and MAFFT identified conserved regions, further examined through entropy-based conservation metrics. Linear B-cell epitopes were predicted via BepiPred 2.0, while VaxiJen and AllerTOP assessed antigenicity and allergenicity. Structural accessibility and functional relevance were validated using Phyre2, Pfam, CDD, and NetSurfP. Results: We identified six epitopes within Influenza A NP and nine within Influenza B NP that are 100% conserved across diverse human and avian subtypes, including H1N1, H2N2, H3N2, H5N1, H7N9, H10N4, and H13N6. The EHPSA motif (residues 81–85) in Influenza A and conserved C-terminal segments in Influenza B emerged as particularly promising surface-accessible, immunogenic, and non-allergenic. Comparative analysis with patents (CN115838421B, US20210341475A1) supports these epitopes' integration into dual-line LFAs and microsphere assays, offering advantages in cost, scalability, and broad strain detection. Conclusions: These conserved NP epitopes represent strong candidates for next-generation, multiplexed lateral flow diagnostics with high sensitivity and specificity, even amidst antigenic drift. | ||||
| Keywords | ||||
| Influenza A& B; nucleoprotein; conserved epitope; antigenic drift; pandemic preparedness | ||||
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