{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T06:43:27Z","timestamp":1784270607352,"version":"3.55.0"},"reference-count":27,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2020,7,14]],"date-time":"2020-07-14T00:00:00Z","timestamp":1594684800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2020YZ014001"],"award-info":[{"award-number":["2020YZ014001"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2021,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Seasonal influenza is an acute respiratory infection causing around 500\u2009000 global deaths annually. There is an unmet medical need to develop more effective antiviral drugs and vaccines against influenza infection. A rapid, accurate, high\u2010throughput titration assay for influenza virus particles or neutralizing antibodies would be extremely useful in these research fields. However, commonly used methods such as tissue culture infective dose and plaque\u2010forming units (PFU) for virus particle quantification, and the plaque reduction neutralization test (PRNT) for antibody determination are time\u2010consuming, laborious, and have limited accuracy. In this study, we developed an efficient assay based on the enzyme\u2010linked immunospot (ELISPOT) technique for the influenza virus and neutralizing antibody titration. Two broad\u2010spectrum antibodies recognizing the nucleoproteins of influenza A and B viruses were used in the assay to broadly and highly sensitively detect influenza virus\u2010infected cells at 16\u2009hours postinfection. An optimized cell culture medium with no tosyl phenylalanyl chloromethyl ketone trypsin and high dose oseltamivir acid was used to improve quantitation accuracy. This ELISPOT assay displayed a good correlation (R<jats:sup>2<\/jats:sup>\u2009=\u20090.9851) with the PFU assay when used to titrate 30 influenza virus isolates. The assay was also applied to measure influenza\u2010neutralizing antibodies in 40 human sera samples, showing a good correlation (R<jats:sup>2<\/jats:sup>\u2009=\u20090.9965) with the PRNT assay. This ELISPOT titration assay is a rapid, accurate, high\u2010throughput assay for quantification of influenza virus and neutralizing antibodies, and provides a powerful tool for research into and development of drugs and vaccines against influenza.<\/jats:p>","DOI":"10.1002\/jmv.26257","type":"journal-article","created":{"date-parts":[[2020,7,4]],"date-time":"2020-07-04T14:04:34Z","timestamp":1593871474000},"page":"3455-3464","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Establishment of a rapid ELISPOT assay for influenza virus titration and neutralizing antibody detection"],"prefix":"10.1002","volume":"93","author":[{"given":"Guosong","family":"Wang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, School of Public Health Xiamen University 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Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, School of Public Health Xiamen University Xiamen Fujian China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Wu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, School of Public Health Xiamen University Xiamen Fujian China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruiqi","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, School of Public Health Xiamen University Xiamen Fujian 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