Expression and self-assembly of virus-like particles from porcine parvovirus and its application in antibody detection

被引:0
作者
Li, Y. [1 ,2 ]
Wang, Q. [3 ]
Yue, W. [1 ,2 ]
Li, X. [1 ,2 ]
Chen, Y. [1 ,2 ]
Gao, Y. [1 ,2 ]
机构
[1] Beijing Biomed Technol Ctr JoFunHwa Biotechnol Nan, Beijing 102600, Peoples R China
[2] 25 Xiangrui St Daxing Dist, Beijing 102600, Peoples R China
[3] Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Anim Dis Control & Prevent, Harbin 150001, Peoples R China
来源
POLISH JOURNAL OF VETERINARY SCIENCES | 2023年 / 26卷 / 04期
关键词
porcine parvovirus; virus -like particles; indirect ELISA; MULTISYSTEMIC WASTING SYNDROME; LINKED-IMMUNOSORBENT-ASSAY; CIRCOVIRUS; ELISA; PIGS; DISEASE; PROTEIN; REPRODUCTION; COINFECTION; PREVALENCE;
D O I
10.24425/pjvs.2023.148280
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
Porcine parvovirus (PPV) is a major causative agent in reproductive pig disease. The swine industry faces a significant economic and epizootic threat; thus, finding a reliable, quick, and practical way to detect it is essential. In this investigation, recombinant PPV VP2 protein was expressed in the Escherichia coli (E. coli) expression systems. As shown by electron microscopy (TEM), Western blot, and hemagglutination (HA) assays, the recombinant VP2 protein was suc-cessfully assembled into virus-like particles (VLPs) after being expressed and purified. These VLPs had a structure that was similar to that of real PPV viruses and also exhibited HA activity. These VLPs induced high levels of PPV-specific antibody titers in mice after immunization, indi-cating that the VLPs may be beneficial as potential candidate antigens. VLPs were used as the coating antigens for the VLP ELISA, and the PPV VLPs-based ELISA displayed a high sensitivity (99%), specificity (93.0%) and agreement rate (98.3%) compared to HI assay, and the agreement rate of this ELISA was 97.5% compared to a commercial ELISA kit. Within a plate, the coefficient of variation (CV) was 10%, and between ELISA plates, the CV was 15%. Accor-ding to a cross-reactivity assay, the technique was PPV-specific in contrast to other viral illness sera. The PPV VLP indirect-ELISA test for PPV detection in pigs with an inactivated vaccine showed that the PPV-positive rate varied among different sample sources from 88.2 to 89.6%. Our results indicate that this ELISA technique was quick, accurate, and repeatable and may be used for extensive serological research on PPV antibodies in pigs.
引用
收藏
页码:611 / 620
页数:10
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