Establishment of an ELISA Based on a Recombinant Antigenic Protein Containing Multiple Prominent Epitopes for Detection of African Swine Fever Virus Antibodies

Dossêh Jean Apôtre Afayibo; Zhonghui Zhang; Hualin Sun; Jingsheng Fu; Yaru Zhao; Tharheer Oluwashola Amuda; Mengli Wu; Junzheng Du; Guiquan Guan; Qingli Niu; Jifei Yang; Hong Yin

doi:10.3390/microorganisms12050943

Microorganisms (May 2024)

Establishment of an ELISA Based on a Recombinant Antigenic Protein Containing Multiple Prominent Epitopes for Detection of African Swine Fever Virus Antibodies

Dossêh Jean Apôtre Afayibo,
Zhonghui Zhang,
Hualin Sun,
Jingsheng Fu,
Yaru Zhao,
Tharheer Oluwashola Amuda,
Mengli Wu,
Junzheng Du,
Guiquan Guan,
Qingli Niu,
Jifei Yang,
Hong Yin

Affiliations

Dossêh Jean Apôtre Afayibo: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Zhonghui Zhang: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Hualin Sun: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Jingsheng Fu: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Yaru Zhao: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Tharheer Oluwashola Amuda: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Mengli Wu: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Junzheng Du: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Guiquan Guan: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Qingli Niu: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Jifei Yang: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China
Hong Yin: State Key Laboratory for Animal Disease Control and Prevention, African Swine Fever Regional Laboratory of China (Lanzhou), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China

DOI: https://doi.org/10.3390/microorganisms12050943
Journal volume & issue: Vol. 12, no. 5
p. 943

Abstract

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African swine fever virus (ASFV) poses a significant threat to the global pig industry, necessitating accurate and efficient diagnostic methods for its infection. Previous studies have often focused on a limited number of epitopes from a few proteins for detecting antibodies against ASFV. Therefore, the current study aimed to use multiple B-cell epitopes in developing an indirect Enzyme-Linked Immunosorbent Assay (ELISA) for enhanced detection of ASFV antibodies. For the expression of recombinant protein, k3 derived from 27 multiple peptides of 11 ASFV proteins, such as p72, pA104R, pB602L, p12, p14.5, p49, pE248R, p30, p54, pp62, and pp220, was used. To confirm the expression of the recombinant protein, we used the Western blotting analysis. The purified recombinant K3 protein served as the antigen in our study, and we employed the indirect ELISA technique to detect anti-ASFV antibodies. The present finding showed that there was no cross-reactivity with antibodies targeting Foot-and-mouth disease virus (FMDV), Porcine circovirus type 2 (PCV2), Pseudorabies virus (PRV), Porcine reproductive and respiratory syndrome virus (PRRSV), and Classical swine fever virus (CSFV). Moreover, the current finding was sensitive enough to find anti-ASFV in serum samples that had been diluted up to 32 times. The test (k3-iELISA) showed diagnostic specificity and sensitivity of 98.41% and 97.40%, respectively. Moreover, during the present investigation, we compared the Ingenasa kit and the k3-iELISA to test clinical pig serum, and the results revealed that there was 99.00% agreement between the two tests, showing good detection capability of the k3-iELISA method. Hence, the current finding showed that the ELISA kit we developed can be used for the rapid detection of ASFV antibodies and used as an alternative during serological investigation of ASF in endemic areas.

Published in Microorganisms

ISSN: 2076-2607 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Science: Biology (General)
Website: https://www.mdpi.com/journal/microorganisms

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