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Cytolysin A (ClyA) is a nanopore sensor with an internal constriction of 3.3 nm, enabling the detection of protein conformations at the single-molecule level. In this study, a nanopore-based approach is developed for analysis of the enzymatic activity of NA, which facilitates rapid and highly sensitive diagnosis of IAV. Current blockade analysis of the d-glucose\/d-galactose-binding protein (GBP) trapped within a type I ClyA-AS (ClyA mutant) nanopore reveals that galactose cleaved from sialyl-galactose by NA of the influenza virus can be detected in real time and at the single-molecule level. Our results show that this nanopore sensor can quantitatively measure the activity of NA with 40\u201380-fold higher sensitivity than those previously reported. Furthermore, the inhibition of NA is monitored using small-molecule antiviral drugs, such as zanamivir. Taken together, our results reveal that the ClyA protein nanopore can be a valuable platform for the rapid and sensitive point-of-care diagnosis of influenza and for drug screening against the NA target.<\/jats:p>","DOI":"10.1021\/acs.analchem.0c03399","type":"journal-article","created":{"date-parts":[[2020,10,14]],"date-time":"2020-10-14T09:54:50Z","timestamp":1602669290000},"page":"14303-14308","source":"Crossref","is-referenced-by-count":33,"title":["Probing the Neuraminidase Activity of Influenza Virus\nUsing a Cytolysin A Protein Nanopore"],"prefix":"10.1021","volume":"92","author":[{"given":"Dong-Kyu","family":"Kwak","sequence":"first","affiliation":[{"name":"KRIBB , , ,","place":["Daejeon, Republic of Korea, 34141"]},{"name":"KRIBB School of Bioscience, University of Science and Technology , , ,","place":["Daejeon, Republic of Korea, 34113"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin-Sik","family":"Kim","sequence":"additional","affiliation":[{"name":"KRIBB , , ,","place":["Daejeon, Republic of Korea, 34141"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4771-7861","authenticated-orcid":true,"given":"Mi-Kyung","family":"Lee","sequence":"additional","affiliation":[{"name":"KRIBB , , ,","place":["Daejeon, Republic of Korea, 34141"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8422-6669","authenticated-orcid":true,"given":"Kyoung-Seok","family":"Ryu","sequence":"additional","affiliation":[{"name":"Korea Basic Science Institute , , 162 Yeongudanji-ro , ,","place":[", Ochang-eup, Cheongju-si, Chungcheongbuk-do, Republic of Korea, 28119"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9381-3251","authenticated-orcid":true,"given":"Seung-Wook","family":"Chi","sequence":"additional","affiliation":[{"name":"KRIBB , , ,","place":["Daejeon, Republic of Korea, 34141"]},{"name":"KRIBB School of Bioscience, University of Science and Technology , , ,","place":["Daejeon, Republic of Korea, 34113"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,10,14]]},"reference":[{"issue":"10127","key":"2026081502163155300_cit1","doi-asserted-by":"publisher","first-page":"1285","DOI":"10.1016\/S0140-6736(17)33293-2","article-title":"Estimates\nof global seasonal influenza-associated respiratory mortality: a modelling\nstudy","volume":"391","author":"Iuliano","year":"2018","journal-title":"Lancet"},{"issue":"1","key":"2026081502163155300_cit2","first-page":"52","article-title":"Isolation of a virus\ncausing respiratory disease in horses","volume":"2","author":"Sovinova","year":"1958","journal-title":"Acta\nVirol."},{"issue":"4536","key":"2026081502163155300_cit3","doi-asserted-by":"publisher","first-page":"1129","DOI":"10.1126\/science.7063847","article-title":"Mass mortality of\nharbor seals: pneumonia associated with influenza A virus","volume":"215","author":"Geraci","year":"1982","journal-title":"Science"},{"issue":"3","key":"2026081502163155300_cit4","doi-asserted-by":"publisher","first-page":"349","DOI":"10.1084\/jem.54.3.349","article-title":"Swine Influenza:\nI. 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