{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,4]],"date-time":"2026-11-04T23:45:13Z","timestamp":1793835913177,"version":"4.6.1"},"reference-count":31,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,5,5]],"date-time":"2023-05-05T00:00:00Z","timestamp":1683244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Med."],"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>Influenza B virus (IBV) is highly contagious, spreads rapidly, and causes seasonal epidemic respiratory disease in the human population, especially in immunocompromised people and young children. Clinical manifestations in this high-risk population are often more severe than in immunocompetent hosts and sometimes atypical. Therefore, rapid, and accurate detection of IBV is important.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>An amplified luminescent proximity homogeneous assay linked immunosorbent assay (AlphaLISA) was developed for detection of IBV by optimizing the ratio of IBV antibody-labeled receptor beads, streptavidin-conjugated donor beads and biotinylated IBV antibody, as well as the optimal temperature and time conditions for incubation. Assay sensitivity, specificity and reproducibility were evaluated. A total of 228 throat swab samples and inactivated influenza B virus were tested by AlphaLISA and lateral flow colloidal gold-based immunoassay (LFIA).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>AlphaLISA produced the best results for detection of inactivated influenza B virus when IBV antibody-labeled acceptor beads were 50\u2009\u03bcg\/ mL, streptavidin-conjugated donor beads were 40\u2009\u03bcg\/mL, and biotinylated IBV antibody was 0.5\u2009\u03bcg\/mL at 37\u00b0C for 15\u201310\u2009min. Under these conditions, AlphaLISA had a limit of detection of 0.24\u2009ng\/mL for the detection of influenza B nucleoprotein, did not cross react with other common respiratory viruses, and showed good reproducibility with inter-assay coefficient of variation (CV) and intra-assay CV\u2009&amp;lt;\u20095%. The results of 228 clinical throat swab samples showed good agreement between AlphaLISA and LFIA (Kappa\u2009=\u20090.982), and AlphaLISA showed better sensitivity than LFIA for detecting inactivated influenza B virus.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>AlphaLISA showed higher sensitivity and throughput in the detection of IBV and can be used for IBV diagnosis and epidemic control.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmed.2023.1155551","type":"journal-article","created":{"date-parts":[[2023,5,5]],"date-time":"2023-05-05T05:51:30Z","timestamp":1683265890000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["Development of an AlphaLISA assay for sensitive and accurate detection of influenza B 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