新开发的多肽-ELISA 法成功检测出绵羊体内针对麦地-维斯纳病毒的抗 IgG 抗体。

IF 1.4 3区 农林科学 Q4 IMMUNOLOGY Veterinary immunology and immunopathology Pub Date : 2024-07-10 DOI:10.1016/j.vetimm.2024.110806
Ecem Su Koçkaya , Hüseyin Can , Yalçın Yaman , Çağrı Kandemir , Turgay Taşkın , Muhammet Karakavuk , Aysu Değirmenci Döşkaya , Mert Döşkaya , Erkan Pehlivan , Halit Deniz Şireli , Adnan Yüksel Gürüz , Cemal Ün
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引用次数: 0

摘要

麦地维斯纳病毒(MVV)是一种可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,因此及时诊断对防治该病的并发症非常重要。在这项研究中,我们的目标是开发一种以来自 gag 蛋白的多肽为抗原的 ELISA。为此,我们预测了 gag 蛋白的 B 细胞表位,并与 B 细胞受体进行了对接分析,以筛选出用于 ELISA 的多肽。在将抗原性最高的三个可溶性表位制备成肽后,使用分为 MVV 阳性(n=24)和阴性(n=13)的绵羊血清样本通过 ELISA 方法测定了每种肽的免疫原性。随后,使用上述免疫原性多肽作为抗原,在室内进行酶联免疫吸附试验,调查绵羊(88 只)的 MVV 血清流行率。结果显示,在三种肽中,有两种肽与 MVV 阳性血清样本发生了强烈反应,且在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)。第三种肽也与阳性血清样本发生了反应,但平均吸光度值没有统计学意义,该肽被认为是非免疫原性的(P=0.175)。根据商业试剂盒,免疫原性的两种肽显示出同样高的灵敏度和特异性,分别为 91.60 和 92.80。此外,以 CKQGSKE 和 CRPQGKAGHKG 肽为抗原的肽-ELISA 检测到的 MVV 血清流行率分别为 3.40 % 和 4.5 %。结果表明,这些多肽可成功用于 MVV 的血清学诊断。
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Newly developed peptide-ELISA successfully detected anti-IgG antibodies against Maedi-Visna virus in sheep

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.

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来源期刊
CiteScore
3.40
自引率
5.60%
发文量
79
审稿时长
70 days
期刊介绍: The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease. Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above. The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.
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