Abstract:
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.
摘要:
MaediVisna病毒(MVV)是一种可以感染绵羊的逆转录病毒。目前还没有针对这种病毒的有效疗法或疫苗,及时诊断对于对抗这种疾病的并发症很重要。在这项研究中,我们旨在使用gag蛋白衍生的肽作为抗原来开发ELISA。为此,预测gag蛋白的B细胞表位并进行与B细胞受体的对接分析以选择用于ELISA的肽。三个具有最高抗原性的可溶性表位作为肽产生后,使用绵羊血清样品通过ELISA测定每种肽的免疫原性,所述绵羊血清样品分类为MVV阳性(n=24)和阴性(n=13)。随后,使用上述免疫原性肽作为抗原的内部ELISA用于研究绵羊中的MVV血清阳性率(n=88)。根据结果,在三种肽中,其中两个与MVV阳性血清样本反应强烈,阳性和阴性血清样本中检测到的平均吸光度值具有统计学意义,表明这些肽具有免疫原性(P=0.016和P=0.038)。第三肽也与阳性血清样品反应,但平均吸光度值在统计学上不显著,并且该肽被认为是非免疫原性的(P=0.175)。根据商业试剂盒,免疫原性两种肽显示91.60和92.80的相同高灵敏度和特异性值。此外,使用CKQGSKE和CRPQGKAGHKG肽作为抗原的肽-ELISA检测到的MVV血清阳性率分别为3.40%和4.5%,分别。因此,结果表明,这些肽可以成功地用于MVV的血清学诊断。