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

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Virologica Sinica 2024年第3期39卷 478-489页

作者： Jiahong Zhu Qingyuan Liu Liuya Li Runyu Zhang Yueting Chang Jiakai Zhao Siyu Liu Xinyu Zhao Xu Chen Yani Sun Qin Zhao Department of preventive Veterinary Medicine College of Veterinary MedicineNorthwest A&F UniversityYangling Observing and Experimental Station of National Data Center of Animal HealthMinistry of AgricultureYangling712100China

African swine fever virus(asfv)poses a significant threat to the global swine ***,there are no effective vaccines or treatments available to combat asfv infection in *** primary means of controlling the spread of the disease is through rapid detection and subsequent elimination of infected ***,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 ***,an Ep402R-deleted asfv variant has been developed as a potential live attenuated vaccine candidate ***,it is crucial to develop detection methods that can distinguish wild-type and Ep402R-deleted asfv *** this study,two recombinant asfv-p72 and-CD2v proteins were expressed using a prokaryotic system and used to immunize Bactrian ***,eight nanobodies against asfv-p72 and ten nanobodies against asfv-CD2v were *** 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 *** two cELISAs exhibited high sensitivity,good specificity,repeatability,and *** coincidence rate between the two cELISAs and commercial ELISA kits was 98.6%and 97.6%,***,the two cELISA for detecting antibodies against asfv demonstrated ease of operation,a low cost,and a simple production *** 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.

关键词： African swine fever virus(asfv) asfv-p72 asfv-CD2v Nanobody-HRp Competitive ELISA

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