Nanobodies against African swine fever virus p72 and CD2v proteins as reagents for developing two cELISAs to detect viral antibodies

Jiahong Zhu; Qingyuan Liu; Liuya Li; Runyu Zhang; Yueting Chang; Jiakai Zhao; Siyu Liu; Xinyu Zhao; Xu Chen; Yani Sun; Qin Zhao

doi:10.1016/j.virs.2024.04.002

June 2024

Citation: Jiahong Zhu, Qingyuan Liu, Liuya Li, Runyu Zhang, Yueting Chang, Jiakai Zhao, Siyu Liu, Xinyu Zhao, Xu Chen, Yani Sun, Qin Zhao. Nanobodies against African swine fever virus p72 and CD2v proteins as reagents for developing two cELISAs to detect viral antibodies .VIROLOGICA SINICA, 2024, 39(3) : 478-489. http://dx.doi.org/10.1016/j.virs.2024.04.002

Nanobodies against African swine fever virus p72 and CD2v proteins as reagents for developing two cELISAs to detect viral antibodies

Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling Observing and Experimental Station of National Data Center of Animal Health, Ministry of Agriculture, Yangling, 712100, China

Corresponding author: Yani Sun, sunyani@nwsuaf.edu.cn
Qin Zhao, qinzhao_2004@nwsuaf.edu.cn
Received Date: 11 September 2023
Accepted Date: 01 April 2024
Available online: 06 April 2024

Abstract

African swine fever virus (ASFV) poses a significant threat to the global swine industry. Currently, there are no effective vaccines or treatments available to combat ASFV infection in pigs. The primary means of controlling the spread of the disease is through rapid detection and subsequent elimination of infected pig. Recently, a lower virulent ASFV isolate with a deleted EP402R gene (CD2v-deleted) has been reported in China, which further complicates the control of ASFV infection in pig farms. Furthermore, an EP402R-deleted ASFV variant has been developed as a potential live attenuated vaccine candidate strain. Therefore, it is crucial to develop detection methods that can distinguish wild-type and EP402R-deleted ASFV infections. In this study, two recombinant ASFV-p72 and -CD2v proteins were expressed using a prokaryotic system and used to immunize Bactrian camels. Subsequently, eight nanobodies against ASFV-p72 and ten nanobodies against ASFV-CD2v were screened. Following the production of these nanobodies with horse radish peroxidase (HRP) fusion proteins, the ASFV-p72-Nb2-HRP and ASFV-CD2v-Nb22-HRP fusions were selected for the development of two competitive ELISAs (cELISAs) to detect anti-ASFV antibodies. The two cELISAs exhibited high sensitivity, good specificity, repeatability, and stability. The coincidence rate between the two cELISAs and commercial ELISA kits was 98.6% and 97.6%, respectively. Collectively, the two cELISA for detecting antibodies against ASFV demonstrated ease of operation, a low cost, and a simple production process. The two cELISAs could determine whether pigs were infected with wild-type or CD2v-deleted ASFV, and could play an important role in monitoring ASFV infections in pig farms.
- African swine fever virus(ASFV)
- , ASFV-p72
- , ASFV-CD2v
- , Nanobody-HRP
- , Competitive ELISA

Electronic Supplementary Material

10.1016j.virs.2024.04.002-ESM.docx
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