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Purification of Crimean-Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis

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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
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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