» Articles » PMID: 38588947

Nanobodies Against African Swine Fever Virus P72 and CD2v Proteins As Reagents for Developing Two CELISAs to Detect Viral Antibodies

Overview
Journal Virol Sin
Specialty Microbiology
Date 2024 Apr 8
PMID 38588947
Authors
Affiliations
Soon will be listed here.
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.

Citing Articles

Development of a nanobody-based competitive enzyme-linked immunosorbent assay for the sensitive detection of antibodies against porcine deltacoronavirus.

Yu R, Zhang L, Bai Y, Zhou P, Yang J, Wang D J Clin Microbiol. 2025; 63(3):e0161524.

PMID: 39950715 PMC: 11898664. DOI: 10.1128/jcm.01615-24.


Identification of a New Conserved Antigenic Epitope by Specific Monoclonal Antibodies Targeting the African Swine Fever Virus Capsid Protein p17.

Xia N, Cao Q, Liu A, Zhang J, Han H, Jiao J Vet Sci. 2024; 11(12).

PMID: 39728990 PMC: 11680328. DOI: 10.3390/vetsci11120650.


A triple protein-based ELISA for differential detection of ASFV antibodies.

Zhang S, Zuo Y, Gu W, Zhao Y, Liu Y, Fan J Front Vet Sci. 2024; 11:1489483.

PMID: 39723184 PMC: 11669292. DOI: 10.3389/fvets.2024.1489483.

References
1.
Caixia W, Songyin Q, Ying X, Haoyang Y, Haoxuan L, Shaoqiang W . Development of a Blocking ELISA Kit for Detection of ASFV Antibody Based on a Monoclonal Antibody Against Full-Length p72. J AOAC Int. 2022; 105(5):1428-1436. DOI: 10.1093/jaoacint/qsac050. View

2.
King D, Reid S, Hutchings G, Grierson S, Wilkinson P, Dixon L . Development of a TaqMan PCR assay with internal amplification control for the detection of African swine fever virus. J Virol Methods. 2002; 107(1):53-61. DOI: 10.1016/s0166-0934(02)00189-1. View

3.
Zhao H, Ren J, Wu S, Guo H, Du Y, Wan B . HRP-conjugated-nanobody-based cELISA for rapid and sensitive clinical detection of ASFV antibodies. Appl Microbiol Biotechnol. 2022; 106(11):4269-4285. PMC: 9130055. DOI: 10.1007/s00253-022-11981-4. View

4.
Cao Y, Han D, Zhang Y, Zhang K, Du N, Tong W . Identification of one novel epitope targeting p54 protein of African swine fever virus using monoclonal antibody and development of a capable ELISA. Res Vet Sci. 2021; 141:19-25. DOI: 10.1016/j.rvsc.2021.10.008. View

5.
Heimerman M, Murgia M, Wu P, Lowe A, Jia W, Rowland R . Linear epitopes in African swine fever virus p72 recognized by monoclonal antibodies prepared against baculovirus-expressed antigen. J Vet Diagn Invest. 2018; 30(3):406-412. PMC: 6505808. DOI: 10.1177/1040638717753966. View