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间接 ELISA 和抗非洲猪瘟病毒 p17 单克隆抗体的检测揭示了高效的检测和应用前景。

The Indirect ELISA and Monoclonal Antibody against African Swine Fever Virus p17 Revealed Efficient Detection and Application Prospects.

机构信息

Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.

College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China.

出版信息

Viruses. 2022 Dec 23;15(1):50. doi: 10.3390/v15010050.

Abstract

Since 2018, the outbreak and prevalence of the African swine fever virus (ASFV) in China have caused huge economic losses. Less virulent ASFVs emerged in 2020, which led to difficulties and challenges for early diagnosis and control of African swine fever (ASF) in China. An effective method of monitoring ASFV antibodies and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, ASFV p17 was successfully expressed in CHO cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on purified p17 was established and optimized. The monoclonal antibody (mAb) against p17 recognized a conservative linear epitope (TETSPLLSH) and exhibited specific reactivity, which was conducive to the identification of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing p17. The ELISA method efficiently detected clinical ASFV infection and effectively monitored the antibody levels in vivo after recombinant PRRSV live vector virus expressing p17 vaccination. Overall, the determination of the conserved linear epitope of p17 would contribute to the in-depth exploration of the biological function of the ASFV antigen protein. The indirect ELISA method and mAb against ASFV p17 revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF.

摘要

自 2018 年以来,非洲猪瘟病毒(ASFV)在中国的爆发和流行造成了巨大的经济损失。2020 年出现了毒力较弱的 ASFVs,这给中国非洲猪瘟(ASF)的早期诊断和控制带来了困难和挑战。迫切需要一种监测 ASFV 抗体和针对 ASFV 的特异性抗体的有效方法,以促进预防技术的发展。本研究采用悬浮培养系统成功地在 CHO 细胞中表达了 ASFV p17。建立并优化了基于纯化 p17 的间接酶联免疫吸附试验(ELISA)。针对 p17 的单克隆抗体(mAb)识别保守线性表位(TETSPLLSH),并表现出特异性反应,有利于鉴定表达 p17 的重组猪繁殖与呼吸综合征病毒(PRRSV)。ELISA 方法可有效检测临床 ASFV 感染,并在接种表达 p17 的重组 PRRSV 活载体病毒后有效监测体内抗体水平。总之,确定 p17 的保守线性表位将有助于深入探索 ASFV 抗原蛋白的生物学功能。针对 ASFV p17 的间接 ELISA 方法和 mAb 显示出高效的检测能力,具有广阔的应用前景,是 ASF 流行病学监测和疫苗研究的理想选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/59f7/9865993/dcf459a61b96/viruses-15-00050-g001.jpg

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