非洲猪瘟病毒(ASFV)对全球养猪业构成重大威胁,需要对其感染进行准确有效的诊断方法。先前的研究通常集中于来自用于检测抗ASFV抗体的少数蛋白质的有限数量的表位。因此,本研究旨在使用多个B细胞表位来开发用于增强ASFV抗体检测的间接酶联免疫吸附测定(ELISA).对于重组蛋白的表达,k3来源于11种ASFV蛋白的27种多肽,如p72,pA104R,pB602L,p12,p14.5,p49,pE248R,使用p30、p54、pp62和pp220。为了证实重组蛋白的表达,我们用了Western印迹分析.纯化的重组K3蛋白作为我们研究的抗原,我们采用间接ELISA技术检测抗ASFV抗体。目前的发现表明,与针对口蹄疫病毒(FMDV)的抗体没有交叉反应,猪圆环病毒2型(PCV2),伪狂犬病病毒(PRV),猪繁殖与呼吸综合征病毒(PRRSV),和经典猪瘟病毒(CSFV)。此外,目前的发现非常敏感,足以在稀释至32倍的血清样本中发现抗ASFV.检测(k3-iELISA)诊断特异性和敏感性分别为98.41%和97.40%,分别。此外,在本次调查中,我们比较了Ingenasa试剂盒和k3-iELISA来测试临床猪血清,结果显示,两次测试之间有99.00%的一致性,显示k3-iELISA方法的良好检测能力。因此,目前的发现表明,我们开发的ELISA试剂盒可用于快速检测ASFV抗体,并可在流行地区ASF的血清学调查中用作替代方法。
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.