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However, the prevalence of CCHF is not monitored in most of the endemic countries due to limited availability of diagnostic assays and biosafety regulations required for handling infectious CCHFV. In this study, we established a protocol to purify the recombinant CCHFV nucleoprotein (NP), which is antigenically highly conserved among multiple lineages\/clades of CCHFVs and investigated its utility in an enzyme-linked immunosorbent assay (ELISA) to detect CCHFV-specific antibodies. The NP gene was cloned into the pCAGGS mammalian expression plasmid and human embryonic kidney 293\u00a0T cells were transfected with the plasmid. The expressed NP molecule was purified from the cell lysate using cesium-chloride gradient centrifugation. Purified NP was used as the antigen for the ELISA to detect anti-CCHFV IgG. Using the CCHFV NP-based ELISA, we efficiently detected CCHFV-specific IgG in anti-NP rabbit antiserum and CCHFV-infected monkey serum. When compared to the commercially available Blackbox CCHFV IgG ELISA kit, our assay showed equivalent performance in detecting CCHFV-specific IgG in human sera. These results demonstrate the usefulness of our CCHFV NP-based ELISA for seroepidemiological studies.<\/jats:p>","DOI":"10.1038\/s41598-021-81752-0","type":"journal-article","created":{"date-parts":[[2021,1,27]],"date-time":"2021-01-27T11:06:38Z","timestamp":1611745598000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Purification of Crimean\u2013Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis"],"prefix":"10.1038","volume":"11","author":[{"given":"Boniface Pongombo","family":"Lombe","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroko","family":"Miyamoto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takeshi","family":"Saito","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Reiko","family":"Yoshida","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rashid","family":"Manzoor","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahiro","family":"Kajihara","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masayuki","family":"Shimojima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuetsu","family":"Fukushi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shigeru","family":"Morikawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomoki","family":"Yoshikawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takeshi","family":"Kurosu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masayuki","family":"Saijo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qing","family":"Tang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Justin","family":"Masumu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Hawman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heinz","family":"Feldmann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ayato","family":"Takada","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,1,27]]},"reference":[{"key":"81752_CR1","doi-asserted-by":"publisher","first-page":"106","DOI":"10.3390\/v8040106","volume":"8","author":"M Zivcec","year":"2016","unstructured":"Zivcec, M., Scholte, F. E., Spiropoulou, C. F., Spengler, J. R. & Bergeron, E. Molecular insights into Crimean\u2013Congo hemorrhagic fever virus. Viruses 8, 106. https:\/\/doi.org\/10.3390\/v8040106 (2016).","journal-title":"Viruses"},{"key":"81752_CR2","doi-asserted-by":"publisher","first-page":"3337","DOI":"10.1099\/vir.0.81213-0","volume":"86","author":"J Chamberlain","year":"2005","unstructured":"Chamberlain, J. et al. Co-evolutionary patterns of variation in small and large RNA segments of Crimean\u2013Congo hemorrhagic fever virus. J. Gen. Virol 86, 3337\u20133341. https:\/\/doi.org\/10.1099\/vir.0.81213-0 (2005).","journal-title":"J. Gen. Virol"},{"key":"81752_CR3","doi-asserted-by":"publisher","first-page":"503","DOI":"10.1093\/trstmh\/trv050","volume":"109","author":"JP Messina","year":"2015","unstructured":"Messina, J. P. et al. The global distribution of Crimean\u2013Congo hemorrhagic fever. Trans. R. Soc. Trop. Med. 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