{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T07:07:59Z","timestamp":1783840079448,"version":"3.55.0"},"reference-count":37,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T00:00:00Z","timestamp":1765929600000},"content-version":"vor","delay-in-days":16,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T00:00:00Z","timestamp":1764547200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["MedComm"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Poliovirus is characterized by three antigenically distinct serotypes that do not elicit cross\u2010neutralizing antibodies. In the final stages of poliovirus eradication, the gold\u2010standard conventional neutralization test (cNT) for detecting serum neutralizing antibodies (NAbs) is highly restricted due to biosafety concerns. To address this, we developed a high\u2010throughput, tri\u2010color pseudovirus\u2010based neutralization assay (PBNA) for the simultaneous quantification of NAbs against all three poliovirus serotypes. We generated pseudoviruses by co\u2010transfecting cells with P1 plasmids, a replication plasmid, and a T7 RNA polymerase plasmid. By optimizing P1 expression, sensitive cell selection (HEK 293T), and plasmid transfection ratios (3:3:1 for P1, replicon, and T7 plasmids), we produced high\u2010titer pseudoviruses (&gt;29\u2010fold increase in titers). Based on high\u2010titer pseudovirus encoding distinct fluorophores (E2, eGFP, and RFP), the PBNA was established, which was optimized for a 12\u00a0h incubation period, 4\u00a0\u00d7\u00a010\u2074 cells per well, and 1500 TCID\n                    <jats:sub>50<\/jats:sub>\n                    \/mL of pseudovirus. It demonstrated high sensitivity, strong serotype specificity, and excellent reproducibility. Furthermore, the PBNA and cNT exhibited excellent congruency (\n                    <jats:italic>r<\/jats:italic>\n                    &gt; 0.88, all serotypes). The tri\u2010color PBNA provides a safe, rapid, and alternative to the cNT, making it an invaluable tool for large\u2010scale serosurveillance, novel vaccine evaluation, and fundamental virological investigations in the post\u2010eradication era.\n                  <\/jats:p>","DOI":"10.1002\/mco2.70551","type":"journal-article","created":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T13:12:10Z","timestamp":1765977130000},"source":"Crossref","is-referenced-by-count":0,"title":["A Pseudovirus\u2010Based Method for the Simultaneous Quantitation of Neutralization Antibodies Against All Three Poliovirus Serotypes"],"prefix":"10.1002","volume":"6","author":[{"given":"Meiyan","family":"Liu","sequence":"first","affiliation":[{"name":"Chinese Academy of Medical Sciences &amp; Peking Union Medical College  Beijing China"},{"name":"Changping Laboratory  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanling","family":"Yu","sequence":"additional","affiliation":[{"name":"Changping Laboratory  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yadong","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology Chinese Academy of Medical Sciences  Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zexin","family":"Tao","sequence":"additional","affiliation":[{"name":"Shandong Center for Disease Control and Prevention  Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Huang","sequence":"additional","affiliation":[{"name":"Changping Laboratory  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xi","family":"Wu","sequence":"additional","affiliation":[{"name":"Division of HIV\/AIDS and Sexually\u2010transmitted Virus Vaccines Institute for Biological Product Control National Institutes for Food and Drug Control (NIFDC)  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases (NITFID) National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention Beijing China"},{"name":"National Polio Laboratory World Health Organization Polio Reference Laboratory for the Western Pacific Region National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention  Beijing China"},{"name":"National Health Commission Key Laboratory of Laboratory Biosafety National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention  Beijing 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