caa a22 4500 605505136 CHVBK 20210128100620.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-5937-7 doi (NATIONALLICENCE)springer-10.1007/s00253-014-5937-7 Isolation of a Bluetongue virus group-specific monoclonal antibody and application to a diagnostic competitive ELISA [Elektronische Daten] [Q. Xu, E. Sun, T. Yang, Y. Feng, H. Geng, T. Wei, J. Li, L. Sun, J. Sun, N. Xi, Q. Zhang, H. Wang, E. Liu, Z. Bu, D. Wu] The Bluetongue virus (BTV) VP7 protein represents an important group-specific antigen that can serve as a basis for diagnostic tests. Here, we report the generation of a novel BTV group-specific monoclonal antibody (Mab; herein named 4H7) that recognizes a conformational epitope in the VP7 protein. We used a phage-displayed peptide screen and site-directed mutagenesis to define the VP7 amino acid residues that most strongly contribute to the conformational epitope recognized by Mab 4H7. Amino acid residues at positions 175, 185, 186, and 278 of the BTV VP7 protein strongly contributed to Mab 4H7 binding. These key amino acid residues are conserved among all BTV serotypes, whereas related Orbiviruses possess at least one amino acid substitution at these positions. We developed a competitive enzyme-linked immunosorbent assay (c-ELISA) using Mab 4H7 and recombinant BTV VP7 protein to detect serum antibodies against this BTV group-specific VP7 epitope. The c-ELISA was used to screen 833 clinical samples collected from animals in three provinces of China. BTV seroprevalence in the three provinces ranged from 25.42 to 47.45%. This work provides the foundation for a new diagnostic c-ELISA that can be further applied to BTV surveillance activities and informs our understanding of the structure of the BTV VP7 protein. Springer-Verlag Berlin Heidelberg, 2014 Bluetongue virus nationallicence VP7 protein nationallicence Monoclonal antibody nationallicence Conformational epitope nationallicence c-ELISA nationallicence Xu Q. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Sun E. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Yang T. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Feng Y. Department of Basic Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, 150030, Harbin, China aut Geng H. Department of Basic Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, 150030, Harbin, China aut Wei T. College of Veterinary Medicine, Jilin Agricultural University, 130000, Changchun, China aut Li J. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Sun L. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Sun J. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Xi N. College of Veterinary Medicine, Jilin Agricultural University, 130000, Changchun, China aut Zhang Q. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Wang H. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Liu E. College of Veterinary Medicine, Jilin Agricultural University, 130000, Changchun, China aut Bu Z. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Wu D. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 729-739 0175-7598 99:2<729 2015 99 253 https://doi.org/10.1007/s00253-014-5937-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-5937-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xu Q. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Sun E. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Yang T. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Feng Y. Department of Basic Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, 150030, Harbin, China aut NATIONALLICENCE 700 1- Geng H. Department of Basic Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, 150030, Harbin, China aut NATIONALLICENCE 700 1- Wei T. College of Veterinary Medicine, Jilin Agricultural University, 130000, Changchun, China aut NATIONALLICENCE 700 1- Li J. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Sun L. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Sun J. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Xi N. College of Veterinary Medicine, Jilin Agricultural University, 130000, Changchun, China aut NATIONALLICENCE 700 1- Zhang Q. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Wang H. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Liu E. College of Veterinary Medicine, Jilin Agricultural University, 130000, Changchun, China aut NATIONALLICENCE 700 1- Bu Z. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 700 1- Wu D. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001, Harbin, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 729-739 0175-7598 99:2<729 2015 99 253