Abstract
Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne pathogen that causes severe disease in humans. CCHFV is widely distributed in more than 30 countries and distinct regions, which means that it poses a serious threat to human health. The nucleocapsid protein (NP) encoded by the CCHFV S gene is the primary detectable antigen in infected cells, which makes it an important viral antigen and a clinical diagnostic target. In this study, the modified biosynthetic peptide (BSP) method was used to identify the fine epitopes on the N- and C- terminals of NP from the CCHFV YL04057 strain using rabbit antiserum against CCHFV-NP. Nine epitopes were identified: E1a (178NLILNRGG185), E1b (184GGDENP189), E2 (352PLKWGKK358), E3 (363FADDS367), E4 (399NPDDAA404), E5a (447DIVASEHL454), E5b (452EHLLHQSL459), E6 (464SPFQNAY470) and E7 (475NATSANII482). Western blotting analysis showed that each epitope interacted with the positive serum of sheep that had been naturally infected with CCHFV. Amino acid sequence alignment between each epitope and their homologous proteins showed that they were almost 100% conserved among 12 CCHFV sequences from different lineages, except for epitopes E1a, E1b and E2. Three-dimensional structural modeling analysis showed that all identified epitopes were located on the surface of the NP “head” domain. This study identified fine epitopes on the N- and C- terminals of NP, which will increase the understanding of the structure and function of NP, and it could lay the foundation for the design and development of a CCHFV multi-epitope peptide vaccine and detection antigen.
Highlights
Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic disease that was first discovered in the West Crimean Peninsula in 1944
We identified the antigenicity of epitopes on nucleocapsid protein (NP) (237– 305) using the same sera of sheep naturally infected with Crimean-Congo hemorrhagic fever virus (CCHFV) [19]
The truncated NP1 and NP3 segments of CCHFV-NP were expressed using the prokaryotic expression vector pET-32a, and each predicted protein was fused with Trix tag, S tag and His tag [22]
Summary
Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic disease that was first discovered in the West Crimean Peninsula in 1944. It has been reported in more than 30 countries and distinct regions in Central and Southern Africa, Southeast Asia, Southeast Europe and the Middle East, with case fatality rates of up to 30–50% [1,2,3]. In 1965, the first case of CCHF in China was found in Bachu County in Xinjiang [4]. CCHF is an important insect-borne disease in China, and it is mainly distributed in Xinjiang, a western region of China. CCHF is characterized by rapid spread and high mortality. There is no effective preventive vaccine or specific antiviral therapy for CCHFV
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