Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection

被引:4
作者
Li, Yanwei [1 ]
Yu, Pingping [1 ]
Bao, Yaxuan [1 ]
Ren, Yuwen [1 ]
Zhao, Shaowei [1 ]
Zhang, Xuexian [1 ]
机构
[1] Beijing Kemufeng Biopharmaceut Co Ltd, 25 Xiangrui St, Beijing 102600, Peoples R China
关键词
Porcine circovirus 2; Virus-like particles; Indirect ELISA; WASTING SYNDROME; CLINICAL SIGNS; TYPE-2; PIGS; PROTEIN; ELISA; VACCINES; HISTORY; DISEASE; SPAIN;
D O I
10.1186/s12917-023-03648-7
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
BackgroundPorcine circovirus 2 (PCV-2) is one of the pathogens that leads to a growing and persistent threat in pigs. Thus, the development of serological detection methods for PCV-2 is of great necessity for clinical diagnosis as well as epidemiological investigations. This study aimed to establish an indirect enzyme-linked immunosorbent assay (ELISA) to examine antibodies against PCV-2 based on virus-like particles (VLPs).ResultsRecombinant PCV-2 Cap protein was expressed in the baculovirus-insect cells system and PCV-2 VLPs were observed over transmission electron microscopy (TEM). The PCV-2 VLPs were shown to have good immunogenicity in mice and stimulated a high level of PCV-2 antibody titers. Using PCV-2 VLPs as coating antigen, the indirect ELISA can detect PCV-2 antibodies in animals with diagnostic sensitivity and specificity of 98.33% and 93.33% compared to immunofluorescence assay (IFA), respectively. The intra- and inter-assay coefficient variations (CVs) were < 10% in a batch, and < 15% in different batches, indicating good repeatability. There was no cross-reaction of this ELISA with antibodies against other porcine viruses. A total of 170 serum samples collected from different pig farms in China were tested for PCV-2 antibodies, and 151 (88.8%) samples were PCV-2 antibody positive.ConclusionOur findings suggest that this ELISA was rapid, specific, and reproducible and can be used for large-scale serological investigations of PCV-2 antibodies in pigs.
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页数:9
相关论文
共 40 条
[1]   Cost of post-weaning multi-systemic wasting syndrome and porcine circovirus type-2 subclinical infection in England An economic disease model [J].
Alarcon, Pablo ;
Rushton, Jonathan ;
Wieland, Barbara .
PREVENTIVE VETERINARY MEDICINE, 2013, 110 (02) :88-102
[2]   Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe [J].
Allan, GM ;
McNeilly, F ;
Kennedy, S ;
Daft, B ;
Clarke, EG ;
Ellis, JA ;
Haines, DM ;
Meehan, BM ;
Adair, BM .
JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION, 1998, 10 (01) :3-10
[3]   Isolation and characterisation of circoviruses from pigs with wasting syndromes in Spain, Denmark and Northern Ireland [J].
Allan, GM ;
Mc Neilly, F ;
Meehan, BM ;
Kennedy, S ;
Mackie, DP ;
Ellis, JA ;
Clark, EG ;
Espuna, E ;
Saubi, N ;
Riera, P ;
Botner, A ;
Charreyre, CE .
VETERINARY MICROBIOLOGY, 1999, 66 (02) :115-123
[4]   Discovery and evolving history of two genetically related but phenotypically different viruses, porcine circoviruses 1 and 2 [J].
Allan, Gordon ;
Krakowka, Steven ;
Ellis, John ;
Charreyre, Catherine .
VIRUS RESEARCH, 2012, 164 (1-2) :4-9
[5]   A review of porcine circovirus 2-associated syndromes and diseases [J].
Chae, C .
VETERINARY JOURNAL, 2005, 169 (03) :326-336
[6]  
Crowther JR., 2000, Methods in Molecular Biology, V149, P1, DOI DOI 10.1385/1592590497
[7]  
Ellis J, 1998, CAN VET J, V39, P44
[8]   Porcine circovirus type 2 protective epitope densely carried by chimeric papaya ringspot virus-like particles expressed in Escherichia coli as a cost-effective vaccine manufacture alternative [J].
Eugenia Aguilera, Brenda ;
Chavez-Calvillo, Gabriela ;
Elizondo-Quiroga, Darwin ;
Noemi Jimenez-Garcia, Monica ;
Carrillo-Tripp, Mauricio ;
Silva-Rosales, Laura ;
Hernandez-Gutierrez, Rodolfo ;
Gutierrez-Ortega, Abel .
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 2017, 64 (03) :406-414
[9]   Efficient application of a baculovirus-silkworm larvae expression system for obtaining porcine circovirus type 2 virus-like particles for a vaccine [J].
He, Qianhua ;
Cao, Zhenming ;
Wang, Pengwei ;
Lu, Qiuyuan ;
Zheng, Hao ;
Sun, Jingchen .
ARCHIVES OF VIROLOGY, 2020, 165 (10) :2301-2309
[10]   Neutralization Mechanism of a Monoclonal Antibody Targeting a Porcine Circovirus Type 2 Cap Protein Conformational Epitope [J].
Huang, Liping ;
Sun, Zhenzhao ;
Xia, Deli ;
Wei, Yanwu ;
Sun, Encheng ;
Liu, Chunguo ;
Zhu, Hongzhen ;
Bian, Haiqiao ;
Wu, Hongli ;
Feng, Li ;
Wang, Jingfei ;
Liu, Changming .
JOURNAL OF VIROLOGY, 2020, 94 (09)