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K205R specific nanobody-horseradish peroxidase fusions as reagents of competitive ELISA to detect African swine fever virus serum antibodies

文献类型： 外文期刊

作者： Zhang, Angke 1 ; Wu, Shuya 1 ; Duan, Xiaohong 3 ; Zhao, Huijun 1 ; Dong, Haoxin 1 ; Ren, Jiahui 1 ; Zhang, Mingfang 1 ; Li, Jiaji 1 ; Duan, Hong 1 ; Zhang, Gaiping 1 ;

作者机构： 1.Henan Agr Univ, Coll Vet Med, Zhengzhou 450046, Henan, Peoples R China

2.Henan Agr Univ, Coll Vet Med, Int Joint Res Ctr Natl Anim Immunol, Zhengzhou 450046, Henan, Peoples R China

3.Hebei Stn Livestock Improvement, Shijiazhuang 050061, Hebei, Peoples R China

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

关键词： Nanobody-HRP; ASFV; K205R; cELISA; Antibody

期刊名称：BMC VETERINARY RESEARCH （ 影响因子：2.792； 五年影响因子：3.008 ）

ISSN：

年卷期： 2022 年 18 卷 1 期

页码：

收录情况： SCI

摘要： Background African swine fever virus (ASFV) is a highly contagious hemorrhagic disease and often lethal, which has significant economic consequences for the swine industry. Due to lacking of commercial vaccine, the prevention and control of ASF largely depend on early large-scale detection and screening. So far, the commercial ELISA kits have a long operation time and are expensive, making it difficult to achieve large-scale clinical applications. Nanobodies are single-domain antibodies produced by camelid animals, and have unique advantages such as smaller molecular weight, easy genetic engineering modification and low-costing of mass production, thus exhibiting good application prospects. Results The present study developed a new method for detection of ASFV specific antibodies using nanobody-horseradish peroxidase (Nb-HRP) fusion proteins as probe. By using camel immunization, phage library construction and phage display technology, five nanobodies against K205R protein were screened. Then, Nb-HRP fusion proteins were produced using genetic modification technology. Based on the Nb-HRP fusion protein as specific antibodies against K205R protein, a new type of cELISA was established to detect ASFV antibodies in pig serum. The cut-off value of the cELISA was 34.8%, and its sensitivity, specificity, and reproducibility were good. Furthermore, the developed cELISA exhibited 99.3% agreement rate with the commercial available ELISA kit (kappa value = 0.98). Conclusions The developed cELISA method has the advantages of simple operation, rapid and low-costing, and can be used for monitoring of ASFV infection in pigs, thus providing a new method for the prevention and control of ASF.

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