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Immunogenicity Characterization of the Recombinant gI Protein Fragment from Pseudorabies Virus and an Evaluation of Its Diagnostic Use in Pigs

文献类型： 外文期刊

作者： He, Haijuan 1 ; Qi, Baojie 1 ; Yang, Yongbo 1 ; Cui, Xiaowen 3 ; Chen, Tianfeng 1 ; Cai, Xuehui 1 ; An, Tongqing 1 ; Wang, Shujie 1 ;

作者机构： 1.Chinese Acad Agr Sci, Harbin Vet Res Inst, Natl Key Lab Anim Dis Control & Prevent, Harbin 150068, Peoples R China

2.Heilongjiang Acad Agr Sci, Inst Anim Husb, Harbin 150086, Peoples R China

3.Heilongjiang Minzu Coll, Harbin 150066, Peoples R China

4.Heilongjiang Res Ctr Vet Biopharmaceut Technol, Harbin 150068, Peoples R China

5.Heilongjiang Prov Key Lab Vet Immunol, Harbin 150068, Peoples R China

关键词： pseudorabies virus; envelope glycoprotein I; immunogenicity; Ni-NTA chromatography; ELISA

期刊名称：VETERINARY SCIENCES （ 影响因子：2.4； ）

ISSN：

年卷期： 2023 年 10 卷 8 期

页码：

收录情况： SCI

摘要： Serological testing is an important method for the diagnosis of pseudorabies virus (PRV) infection. We aimed to investigate the envelope glycoprotein I (gI) of PRV, a strong immunogen, and its potential as an efficient and low-cost diagnostic reagent. In this study, the DNA of the PRV SC strain was used as the template, and the recombinant fragment of gI (633 bp) was amplified via PCR using synthetic primers, and was then ligated into the pET-30a expression vector. The constructs were transferred into Escherichia coli (E. coli) for prokaryotic expression, and the antigenicity of the expression products was identified byWestern blot analysis with pig positive serum against PRV. The recombinant protein was purified by a Ni column, and BALB/c mice were immunized with purified gI protein to obtain anti-gI-positive serum. After PK-15 cells had been infected by PRV for 48 h, the immunogenicity of purified gI protein was identified with a fluorescence immunoassay using anti-gI mouse serum. The recombinant plasmid (pET-30a-gI) was expressed, and the native gI protein was obtained after denaturation by urea and renaturation by dialysis. A small-scale ELISA test containing 1.0 mu g/mL of purified gI protein was designed to evaluate pig serum (80 samples), and the results of the ELISA test were compared to those of competitive ELISA (cELISA) tests using IDEXX Kits, which resulted in 97.5% consistency. The results suggested that the truncated gI protein may be a potential diagnostic reagent.

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