基于噬菌体纳米体的非洲猪瘟病毒抗体竞争性免疫检测方法。

IF 4 3区 医学 Q2 VIROLOGY
Wenting Chen, Jifei Yang, Qingli Niu, Jinming Wang, Yanghe Liu, Xuesai Li, Yaru Zhao, Zhonghui Zhang, Zhijie Liu, Guiquan Guan, Hong Yin
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引用次数: 0

摘要

非洲猪瘟(ASF)是一种高度传染性和破坏性疾病,对全球养猪业构成重大威胁。非洲猪瘟的迅速传播及其持续的大流行继续影响着全世界的养猪业。由于缺乏有效的疫苗,加上非洲猪瘟病毒(ASFV)的复杂性,使得控制和根除非洲猪瘟成为一项艰巨的挑战。来源于骆驼的纳米抗体作为传统单克隆抗体的替代品,在各种生物应用中具有明显的优势。在这项研究中,从噬菌体展示的免疫文库中选择了靶向ASFV K205R蛋白的特异性纳米体。根据互补决定区(cdr)分离出10个纳米体,其中4个纳米体与ASFV幼稚型K205R蛋白结合。经过评估,选择纳米体vh1用于开发竞争性酶联免疫吸附试验(ELISA),用于ASFV抗体检测。对不同反应条件下的检测结果进行优化,检测截止值为26.85%,诊断敏感性和特异性分别为97.52%和97.48%。该方法与感染其他猪病毒的猪血清无交叉反应,检测灵敏度为1:128。临床样品对比分析显示,基于纳米抗体的elisa与基于单克隆抗体的elisa (Mab-cELISA)具有较高的一致性(98.98%)。综上所述,本研究提出了一种基于噬菌体纳米体的竞争性ELISA检测ASFV抗体,可用于ASF血清监测。此外,这里鉴定的k205r特异性纳米体可能适用于其他生物学或生物医学应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A phage-displayed nanobody-based competitive immunoassay for the detection of African swine fever virus antibodies.

African swine fever (ASF) is a highly infectious and devastating disease that poses a significant threat to the global swine industry. The rapid spread of ASF and its ongoing pandemics continue to impact pig farming worldwide. The absence of an effective vaccine, coupled with the complexity of the African swine fever virus (ASFV), makes the control and eradication of ASF a formidable challenge. Nanobodies, derived from camelids, have emerged as promising alternatives to conventional monoclonal antibodies, offering distinct advantages in various biological applications. In this study, specific nanobodies targeting the ASFV K205R protein were selected from a phage-displayed immune library. Ten individual nanobodies were isolated based on their complementary determining regions (CDRs), and four were found to bind to the naive K205R protein of ASFV. After evaluation, nanobody VHH1 was selected for the development of a competitive enzyme-linked immunosorbent assay (ELISA) for ASFV antibody detection. The assay was optimized for various reaction conditions, and the cut-off value was determined to be 26.85%, with diagnostic sensitivity and specificity of 97.52% and 97.48%, respectively. No cross-reactivity was observed with sera from pigs infected with other swine viruses, and the assay exhibited a detection sensitivity of 1:128. Comparative analysis of clinical samples showed a high concordance rate (98.98%) between the nanobody-based and monoclonal antibody-based ELISAs (Mab-cELISA). In conclusion, this study presents a phage-displayed nanobody-based competitive ELISA for the detection of ASFV antibodies, which could be valuable for ASF sero-surveillance. Additionally, the K205R-specific nanobodies identified here may be adapted for other biological or biomedical applications.

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来源期刊
Virology Journal
Virology Journal 医学-病毒学
CiteScore
7.40
自引率
2.10%
发文量
186
审稿时长
1 months
期刊介绍: Virology Journal is an open access, peer reviewed journal that considers articles on all aspects of virology, including research on the viruses of animals, plants and microbes. The journal welcomes basic research as well as pre-clinical and clinical studies of novel diagnostic tools, vaccines and anti-viral therapies. The Editorial policy of Virology Journal is to publish all research which is assessed by peer reviewers to be a coherent and sound addition to the scientific literature, and puts less emphasis on interest levels or perceived impact.
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