一株猪圆环病毒2d基因型毒株的全基因组序列及其衣壳蛋白的小鼠免疫原性分析

doi:10.11843/j.issn.0366-6964.2023.10.025

Abstract

Abstract: The main objectives of this study were to investigate the main pathogens causing postweaning multisystemic wasting syndrome (PMWS) on a large-scale pig farm in Hubei Province. In this study, tissue samples were collected from piglets with typical clinical symptoms of PMWS in this pig farm, and PCR detection revealed the presence of porcine circovirus type 2 (PCV2) infection. The complete genome of PCV2 was cloned and sequenced. Then, the sequence of the complete genome and key genes of the positive samples were compared with other PCV2 strains around the world, and their nucleotide homology was analyzed. The virus strain was isolated, cultured, and identified by using porcine kidney epithelial cell line PK-15, and the epitope and immunogenicity in mice of nucleocapsid protein Cap were analyzed and evaluated. The isolation and identification results showed that we successfully isolated a PCV2 strain (name:XY2022; GenBank accession No.:OM249965.1). The complete genome length of XY2022 was 1 767 bp, and the titer of XY2022 on isolated cells was 10^5.70TCID₅₀·mL^-1. Sequence analysis showed that XY2022 belonged to PCV2d, the nucleotide homology of the reference strains was 94.3%-99.8%, and that of ORF2 was 89.2%-99.6%. The results of immunogenicity analysis and evaluation of Cap showed that XY2022 and other genotypes of PCV2 had different sites in the linear immunodominant region, linear neutralization epitope, and conformational neutralization epitope. Both PCV2d (XY2022) Cap and PCV2b (WH) Cap can induce the production of neutralizing antibodies to inhibit PCV2-infection in mice, but the level of neutralizing antibody induced by 2d Cap is more stable. This study provides a reference for the epidemiological study of PCV2 and the prevention and control of the mutated strains in Hubei Province and provides materials for the study of the pathogenesis of PCV2 and the development of novel vaccines.

Key words: porcine circovirus type 2, virus isolation and identification, sequence analysis, immunogenicity

CLC Number:

S852.659.2

LI Chang, ZHOU Ping, WEN Ping, YANG Keli, LIU Wei, GUO Rui, LIU Zewen, YUAN Fangyan, GAO Ting, SONG Haofei, TIAN Yongxiang, ZHOU Danna. Investigation of the Genome-wide Sequence and Immunogenicity of Capsid Protein in Mice for a Porcine Circovirus 2d Genotype Strain[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4289-4299.

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Investigation of the Genome-wide Sequence and Immunogenicity of Capsid Protein in Mice for a Porcine Circovirus 2d Genotype Strain

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