{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T12:14:11Z","timestamp":1784808851900,"version":"3.55.0"},"reference-count":26,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2014,5,1]],"date-time":"2014-05-01T00:00:00Z","timestamp":1398902400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2014,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>To assess the efficacy of novel antiviral drugs against influenza virus in clinical trials, it is necessary to quantify infectious virus titers in respiratory tract samples from patients. Typically, this is achieved by inoculating virus-susceptible cells with serial dilutions of clinical specimens and detecting the production of progeny virus by hemagglutination, since influenza viruses generally have the capacity to bind and agglutinate erythrocytes of various species through their hemagglutinin (HA). This readout method is no longer adequate, since an increasing number of currently circulating influenza A virus H3 subtype (A[H3]) viruses display a reduced capacity to agglutinate erythrocytes. Here, we report the magnitude of this problem by analyzing the frequency of HA-deficient A(H3) viruses detected in The Netherlands from 1999 to 2012. Furthermore, we report the development and validation of an alternative method for monitoring the production of progeny influenza virus in quantitative virus cultures, which is independent of the capacity to agglutinate erythrocytes. This method is based on the detection of viral nucleoprotein (NP) in virus culture plates by enzyme-linked immunosorbent assay (ELISA), and it produced results similar to those of the hemagglutination assay using strains with good HA activity, including A\/Brisbane\/059\/07 (H1N1), A\/Victoria\/210\/09 (H3N2), other seasonal A(H1N1), A(H1N1)pdm09, and the majority of A(H3) virus strains isolated in 2009. In contrast, many A(H3) viruses that have circulated since 2010 failed to display HA activity, and infectious virus titers were determined only by detecting NP. The virus culture ELISA described here will enable efficacy testing of new antiviral compounds in clinical trials during seasons in which nonhemagglutinating influenza A viruses circulate.<\/jats:p>","DOI":"10.1128\/jcm.03575-13","type":"journal-article","created":{"date-parts":[[2014,3,13]],"date-time":"2014-03-13T01:02:55Z","timestamp":1394672575000},"page":"1672-1677","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":35,"title":["Detection of Nonhemagglutinating Influenza A(H3) Viruses by Enzyme-Linked Immunosorbent Assay in Quantitative Influenza Virus Culture"],"prefix":"10.1128","volume":"52","author":[{"given":"C. A.","family":"van Baalen","sequence":"first","affiliation":[{"name":"Viroclinics Biosciences B.V., Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C.","family":"Els","sequence":"additional","affiliation":[{"name":"Viroclinics Biosciences B.V., Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L.","family":"Sprong","sequence":"additional","affiliation":[{"name":"Viroclinics Biosciences B.V., Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"van Beek","sequence":"additional","affiliation":[{"name":"Erasmus MC Department of Viroscience, Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"van der Vries","sequence":"additional","affiliation":[{"name":"Erasmus MC Department of Viroscience, Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A. D. M. E.","family":"Osterhaus","sequence":"additional","affiliation":[{"name":"Viroclinics Biosciences B.V., Rotterdam, The Netherlands"},{"name":"Erasmus MC Department of Viroscience, Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G. F.","family":"Rimmelzwaan","sequence":"additional","affiliation":[{"name":"Viroclinics Biosciences B.V., Rotterdam, The Netherlands"},{"name":"Erasmus MC Department of Viroscience, Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2005.08.006"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1218841110"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0066325"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1186\/1743-422X-4-42"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2001.1121"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2000.0679"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00398-08"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.37.4.937-943.1999"},{"key":"e_1_3_3_10_2","volume-title":"Manual for the laboratory diagnosis and virological surveillance of influenza","author":"WHO","year":"2011","unstructured":"WHO. 2011. Manual for the laboratory diagnosis and virological surveillance of influenza. WHO Press, Geneva, Switzerland. http:\/\/whqlibdoc.who.int\/publications\/2011\/9789241548090_eng.pdf."},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-0934(98)00071-8"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmoldx.2012.11.007"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01863914"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01310622"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2011.11.006"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2009.11.043"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1586\/eri.11.51"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1244730"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01023-13"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1097211"},{"key":"e_1_3_3_21_2","doi-asserted-by":"crossref","first-page":"1814","DOI":"10.4049\/jimmunol.126.5.1814","article-title":"Expression of influenza A virus internal antigens on the surface of infected P815 cells","volume":"126","author":"Yewdell JW","year":"1981","unstructured":"YewdellJWFrankEGerhardW. 1981. Expression of influenza A virus internal antigens on the surface of infected P815 cells. J. Immunol. 126:1814\u20131819.","journal-title":"J. Immunol."},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2013.06.071"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.28.6.1308-1313.1990"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-73-10-2737"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00229-11"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2009.01.116"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-540-92165-3_21"}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.03575-13","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.03575-13","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,11]],"date-time":"2023-07-11T04:48:28Z","timestamp":1689050908000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JCM.03575-13"}},"subtitle":[],"editor":[{"given":"A. J.","family":"McAdam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2014,5]]},"references-count":26,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2014,5]]}},"alternative-id":["10.1128\/JCM.03575-13"],"URL":"https:\/\/doi.org\/10.1128\/jcm.03575-13","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,5]]},"assertion":[{"value":"2013-12-23","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-03-04","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-04-17","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}