TY - JOUR
T1 - Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis
AU - Lombe, Boniface Pongombo
AU - Miyamoto, Hiroko
AU - Saito, Takeshi
AU - Yoshida, Reiko
AU - Manzoor, Rashid
AU - Kajihara, Masahiro
AU - Shimojima, Masayuki
AU - Fukushi, Shuetsu
AU - Morikawa, Shigeru
AU - Yoshikawa, Tomoki
AU - Kurosu, Takeshi
AU - Saijo, Masayuki
AU - Tang, Qing
AU - Masumu, Justin
AU - Hawman, David
AU - Feldmann, Heinz
AU - Takada, Ayato
N1 - Funding Information:
We thank A. Mori-Kajihara for technical assistance, K. Ibaraki for administrative assistance, K. Barrymore for editing the manuscript, and all members of our laboratory for helpful discussions and admirable technical assistance. This work was supported by the Agency for Medical Research and Development (AMED) and Japan International Cooperation Agency (JICA) within the framework of the Science and Technology Research Partnership for Sustainable Development (SATREPS) (JP18jm0110019), partly by the Japanese Initiative for Progress of Research on Infectious Disease for Global Epidemics (J-PRIDE) from AMED (JP19fm0208101), a research grant from the Ministry of Health, Labour, and Welfare of Japan (H29-Sinkogyousei-Shitei-002), and the Intramural Research Program of the NIAID, NIH.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - 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.
AB - 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.
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U2 - 10.1038/s41598-021-81752-0
DO - 10.1038/s41598-021-81752-0
M3 - Article
C2 - 33504869
AN - SCOPUS:85099844153
SN - 2045-2322
VL - 11
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 2324
ER -