Ege University Faculty of Science Department of Biology Molecular Biology Section, İzmir, Türkiye.
Ege University Faculty of Science Department of Biology Molecular Biology Section, İzmir, Türkiye; Ege University Vaccine Development Application and Research Center, İzmir, Türkiye.
Vet Immunol Immunopathol. 2024 Aug;274:110806. doi: 10.1016/j.vetimm.2024.110806. Epub 2024 Jul 10.
Maedi Visna Virus (MVV) is a retrovirus that can infect sheep. There is still no effective therapy or vaccine against this virus and timely diagnosis is important to combat the complications of the disease. In this study, we aimed to develop an ELISA using peptides derived from gag protein as antigen. For this purpose, B cell epitopes of gag protein were predicted and a docking analysis with the B cell receptor was performed to select peptides to be used in ELISA. After three soluble epitopes with the highest antigenicity were produced as peptides, the immunogenicity of each peptide was determined by ELISA using sheep serum samples categorized as MVV positive (n=24) and negative (n=13). Subsequently, in house ELISA using above mentioned immunogenic peptides as antigen was used to investigate MVV seroprevalence in sheep (n=88). According to the results, among three peptides, two of them strongly reacted with MVV positive serum samples and the mean absorbance values detected among positive and negative serum samples were statistically significant, indicating that these peptides were immunogenic (P=0.016 and P=0.038). The third peptide also reacted with positive serum samples but the mean absorbance value was not statistically significant and this peptide was considered non-immunogenic (P=0.175). The immunogenic two peptides showed the same high sensitivity and specificity values of 91.60 and 92.80 according to the commercial kit. Moreover, MVV seroprevalence detected by peptide-ELISAs using CKQGSKE and CRPQGKAGHKG peptides as antigen was 3.40 % and 4.5 %, respectively. As a result, it was shown that these peptides can be successfully used for serological diagnosis of MVV.
绵羊梅迪-维斯纳病毒（MVV）是一种能够感染绵羊的逆转录病毒。目前针对该病毒尚无有效的治疗方法或疫苗，因此及时诊断对于防治该疾病的并发症非常重要。在本研究中，我们旨在使用来自 gag 蛋白的肽段作为抗原开发 ELISA。为此，预测了 gag 蛋白的 B 细胞表位，并进行了 B 细胞受体对接分析，以选择用于 ELISA 的肽段。在生成三个可溶性抗原性最高的肽段作为肽段后，使用归类为 MVV 阳性（n=24）和阴性（n=13）的绵羊血清样本通过 ELISA 确定了每种肽段的免疫原性。随后，使用上述免疫原性肽段作为抗原的内部 ELISA 用于研究绵羊中的 MVV 血清流行率（n=88）。结果表明，在三种肽段中，有两种与 MVV 阳性血清样本强烈反应，阳性和阴性血清样本之间检测到的平均吸光度值具有统计学意义，表明这些肽段具有免疫原性（P=0.016 和 P=0.038）。第三种肽段也与阳性血清样本反应，但平均吸光度值无统计学意义，被认为是非免疫原性的（P=0.175）。根据商业试剂盒，两种免疫原性肽段的灵敏度和特异性值均为 91.60%和 92.80%。此外，使用 CKQGSKE 和 CRPQGKAGHKG 肽段作为抗原的肽 ELISA 检测到的 MVV 血清流行率分别为 3.40%和 4.5%。结果表明，这些肽段可成功用于 MVV 的血清学诊断。
