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Establishment and characterization of a novel indirect ELISA method based on ASFV antigenic epitope-associated recombinant protein

文献类型： 外文期刊

作者： Jin, Jiaxin 1 ; Bai, Yilin 3 ; Zhang, Yuanyuan 1 ; Lu, Wenlong 1 ; Zhang, Shuai 1 ; Zhao, Xuyang 1 ; Sun, Yaning 4 ; Wu, Yanan 1 ; Zhang, Angke 1 ; Zhang, Gaiping 1 ; Sun, Aijun 1 ; Zhuang, Guoqing 1 ;

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

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

3.Zhengzhou Univ, Sch Agr Sci, Zhengzhou, Henan, Peoples R China

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

5.Henan Acad Agr Sci, Henan Prov Key Lab Anim Immunol, Zhengzhou, Peoples R China

6.Longhu Lab Adv Immunol, Zhengzhou, Peoples R China

关键词： African swine fever; African swine fever virus; Antigenic epitope; Recombinant protein; Indirect ELISA

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.2； 五年影响因子：7.8 ）

ISSN： 0141-8130

年卷期： 2023 年 253 卷

页码：

收录情况： SCI

摘要： African Swine Fever (ASF) is an acute and highly lethal disease in pigs caused by African Swine Fever Virus (ASFV). Viral proteins have been commonly used as antigenic targets for the development of ASF diagnostic methods. However, the prokaryotic expression of viral proteins has deficiencies such as instability, insolubility, and high cost in eukaryotic situations. This study screened and verified ASFV-encoded p72, p54, and p30 protein antigenic epitopes. Subsequently, a novel antigenic epitope-associated recombinant protein was designed based on an ideal structural protein and expressed in Escherichia coli (E. coli). Western blot analysis indicated that the recombinant protein could specifically react with the monoclonal antibody (mAb) of p72 and polyclonal anti-bodies of p54 and p30, respectively. Next, an ASF indirect ELISA (iELISA) method was established based on the recombinant protein, which has no specific reaction with sera of other important pig viral diseases. Meanwhile, it shows a sensitivity to detecting dilutions of ASF-positive reference serum up to 1:6400. The clinical sample detection results showed a high coincidence rate of 98 % with a commercial competition ELISA kit. In conclusion, we established a novel specific, and sensitive ASF serologic detection method that opens new avenues for ASF serodiagnostic method development.

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