Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis
Abstract Crimean–Congo hemorrhagic fever virus (CCHFV) causes a zoonotic disease, Crimean–Congo hemorrhagic fever (CCHF) endemic in Africa, Asia, the Middle East, and Southeastern Europe. However, the prevalence of CCHF is not monitored in most of the endemic countries due to limited availability of...
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2021
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oai:doaj.org-article:990474cfe0c84f5c9b92d8fc56b092032021-12-02T13:23:58ZPurification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis10.1038/s41598-021-81752-02045-2322https://doaj.org/article/990474cfe0c84f5c9b92d8fc56b092032021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81752-0https://doaj.org/toc/2045-2322Abstract Crimean–Congo hemorrhagic fever virus (CCHFV) causes a zoonotic disease, Crimean–Congo hemorrhagic fever (CCHF) endemic in Africa, Asia, the Middle East, and Southeastern Europe. 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 T 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.Boniface Pongombo LombeHiroko MiyamotoTakeshi SaitoReiko YoshidaRashid ManzoorMasahiro KajiharaMasayuki ShimojimaShuetsu FukushiShigeru MorikawaTomoki YoshikawaTakeshi KurosuMasayuki SaijoQing TangJustin MasumuDavid HawmanHeinz FeldmannAyato TakadaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021) |
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Medicine R Science Q Boniface Pongombo Lombe Hiroko Miyamoto Takeshi Saito Reiko Yoshida Rashid Manzoor Masahiro Kajihara Masayuki Shimojima Shuetsu Fukushi Shigeru Morikawa Tomoki Yoshikawa Takeshi Kurosu Masayuki Saijo Qing Tang Justin Masumu David Hawman Heinz Feldmann Ayato Takada Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
| description |
Abstract Crimean–Congo hemorrhagic fever virus (CCHFV) causes a zoonotic disease, Crimean–Congo hemorrhagic fever (CCHF) endemic in Africa, Asia, the Middle East, and Southeastern Europe. 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 T 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. |
| format |
article |
| author |
Boniface Pongombo Lombe Hiroko Miyamoto Takeshi Saito Reiko Yoshida Rashid Manzoor Masahiro Kajihara Masayuki Shimojima Shuetsu Fukushi Shigeru Morikawa Tomoki Yoshikawa Takeshi Kurosu Masayuki Saijo Qing Tang Justin Masumu David Hawman Heinz Feldmann Ayato Takada |
| author_facet |
Boniface Pongombo Lombe Hiroko Miyamoto Takeshi Saito Reiko Yoshida Rashid Manzoor Masahiro Kajihara Masayuki Shimojima Shuetsu Fukushi Shigeru Morikawa Tomoki Yoshikawa Takeshi Kurosu Masayuki Saijo Qing Tang Justin Masumu David Hawman Heinz Feldmann Ayato Takada |
| author_sort |
Boniface Pongombo Lombe |
| title |
Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
| title_short |
Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
| title_full |
Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
| title_fullStr |
Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
| title_full_unstemmed |
Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
| title_sort |
purification of crimean–congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
| publisher |
Nature Portfolio |
| publishDate |
2021 |
| url |
https://doaj.org/article/990474cfe0c84f5c9b92d8fc56b09203 |
| work_keys_str_mv |
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| _version_ |
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