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Development of an Indirect ELISA Based on VP1-CRT Fusion Protein for Detection of FMDV-O in Swine

文献类型： 外文期刊

作者： Liu, Chang 1 ; Feng, Hua 2 ; Liu, Yunchao 2 ; Chen, Yumei 3 ; Yang, Suzhen 2 ; Lu, Qingxia 2 ; Feng, Lili 4 ; Deng, Ruigu 1 ;

作者机构： 1.Jilin Univ, Coll Vet Med, Changchun 130033, Jilin, Peoples R China

2.Henan Acad OfAgr Sci, Henan Prov Key Lab Anim Immunol, Minist Agr, Key Lab Anim Immunol, Zhengzhou 450002, Henan, Peoples R China

3.Zhengzhou Univ, Sch Life Sci, Zhengzhou 450001, Henan, Peoples R China

4.Henan Acad Agr Sci, Inst Agr Econ & Informat, Zhengzhou 450002, Peoples R China

5.Henan Agr Univ, Coll Anim Sci & Vet Med, Zhengzhou 450002, Henan, Peoples R China

关键词： FMDV-O VP1; Fusion protein; Polymers; Prokaryotic expression; Indirect ELISA

期刊名称：INTERNATIONAL JOURNAL OF AGRICULTURE AND BIOLOGY （ 影响因子：0.822； 五年影响因子：0.906 ）

ISSN： 1560-8530

年卷期： 2020 年 23 卷 1 期

页码：

收录情况： SCI

摘要： Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals The open reading frame (ORF) of VP1 codes the most important structure protein of foot and mouth disease virus (FMDV). To develop a highly sensitive and specific indirect ELISA method for detecting FMDV-O antibody, the prokaryotic optimized gene vp1 was fused with truncated calceticulin (rT5V) and co-expressed with trigger factor (Tf16) which could assist soluble expression of antigenicity polymers in Escherichia coli. The rT5V was purified using size-exclusion chromatography with the purity about 80%. The rT5V-ELISA method was developed using the purified rT5V as coating antigen for FMDV-O antibody detection. The sensitivity and specificity of the rT5V-ELISA were 84.2 and 100%, respectively. The comparative test between rT5V-ELISA and the liquid-phase blocking ELISA kit (LPB-ELISA) with 376 clinical samples showed the coincidence rate was 84.04%. In conclusion, the rT5V was achieved in soluble polymers by co-expressing with Tf16 in E. coli. The rT5V-ELISA was developed as a highly sensitive and specific method for FMDV-O antibody monitoring after infection and/or vaccination. (C) 2020 Friends Science Publishers

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