Title: | Purification of Crimean-Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis |
Authors: | Lombe, Boniface Pongombo Browse this author |
Miyamoto, Hiroko Browse this author |
Saito, Takeshi Browse this author |
Yoshida, Reiko Browse this author →KAKEN DB |
Manzoor, Rashid Browse this author |
Kajihara, Masahiro Browse this author →KAKEN DB |
Shimojima, Masayuki Browse this author →KAKEN DB |
Fukushi, Shuetsu Browse this author →KAKEN DB |
Morikawa, Shigeru Browse this author →KAKEN DB |
Yoshikawa, Tomoki Browse this author →KAKEN DB |
Kurosu, Takeshi Browse this author →KAKEN DB |
Saijo, Masayuki Browse this author →KAKEN DB |
Tang, Qing Browse this author |
Masumu, Justin Browse this author |
Hawman, David Browse this author |
Feldmann, Heinz Browse this author |
Takada, Ayato Browse this author →KAKEN DB |
Issue Date: | 27-Jan-2021 |
Publisher: | Nature Research |
Journal Title: | Scientific reports |
Volume: | 11 |
Issue: | 1 |
Start Page: | 2324 |
Publisher DOI: | 10.1038/s41598-021-81752-0 |
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. |
Type: | article |
URI: | http://hdl.handle.net/2115/82393 |
Appears in Collections: | 人獣共通感染症国際共同研究所 (International Institute for Zoonosis Control) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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