The development of a multiplexed Soluble Phage Array (SPAr) for the detection of zoonotic pathogens

开发用于检测人畜共患病病原体的多重可溶性噬菌体阵列 (SPAr)

• 批准号: BB/V016148/1
• 负责人: Kevin Gough
• 金额: $ 19.15万
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV016148%2F1
• 关键词: development; multiplexed; Soluble; Phage; Array

Serosurveillance is a valuable approach to disease monitoring in animal populations to facilitate effective intervention and containment strategies. It has been suggested that effective serosurveillance of zoonotic diseases could provide an early-warning system for the potential emergence of a zoonotic pathogen in human populations. However, to date, serosurveillance is somewhat limited by a single-pathogen focus and lacks the ability to distinguish between very closely related pathogens. Therefore, there is a need for an affordable, accessible, multiplexed approach with very high specificity to monitor for a wide range of established and emerging pathogens. The huge diversity of phage display peptide libraries combined with the screening power of next generation sequencing (NGS) in a process termed next generation phage display (NGPD) can be applied to discover large panels of peptide mimotopes recognised by antibody responses to disease. We propose that a phage-peptide sub-library enriched to contain multiple infection-specific mimotopes can itself be used as the assay 'antigen' to diagnose infection with a particular pathogen; an assay format we have termed Soluble Phage Array (SPAr). This assay would measure recognition of the panel of infection-specific phage-displayed mimotopes by antibodies in serum samples by NGS analysis of phage that are bound by the antibodies. We hypothesise that NGPD can be used to develop SPAr assays that can accurately identify infection with zoonotic pathogens and that these can be multiplexed with high accuracy and reproducibility.This proof-of-concept study will focus on four emergent zoonotic viruses that affect sheep or horses. These are louping ill (LIV), tick-borne encephalitis (TBEV) and Rift Valley fever (RVFV) viruses in sheep, and West Nile virus (WNV) in horses. The selected viruses represent the genera Flavivirus (LIV, TBEV and WNV) and Phlebovirus (RVFV). These viruses have been carefully chosen to represent the complexity of serosurveillance of emerging viral zoonoses in order to demonstrate the potential of the SPAr assay. A multiplexed SPAr assay would have considerable advantages over existing serosurveillance approaches. It would not be limited to a single-pathogen approach and would have the ability to distinguish very closely related pathogens, a property vital to inform effective control strategies for zoonotic pathogens.

血清监测是动物群体疾病监测的一种有价值的方法，可促进有效的干预和遏制策略。有人建议，对人畜共患疾病进行有效的血清监测可以为人群中可能出现的人畜共患病原体提供预警系统。然而，迄今为止，血清监测在某种程度上受到单一病原体焦点的限制，并且缺乏区分非常密切相关的病原体的能力。因此，需要一种负担得起、易于获得、具有极高特异性的多重方法来监测各种已确定的和新出现的病原体。噬菌体展示肽文库的巨大多样性与下一代噬菌体展示 (NGPD) 过程中下一代测序 (NGS) 的筛选能力相结合，可用于发现疾病抗体反应所识别的大量肽模拟表位。我们建议，富含多种感染特异性模拟表位的噬菌体肽子库本身可以用作检测“抗原”来诊断特定病原体的感染；我们将这种检测格式称为可溶性噬菌体阵列 (SPAr)。该测定将通过对抗体结合的噬菌体进行 NGS 分析来测量血清样品中的抗体对一组感染特异性噬菌体展示的模拟表位的识别。我们假设 NGPD 可用于开发 SPAr 检测，能够准确识别人畜共患病原体的感染，并且这些检测可以高精度和可重复性地进行多重检测。这项概念验证研究将重点关注影响羊或马的四种新出现的人畜共患病毒。这些病毒包括绵羊中的放大病 (LIV)、蜱传脑炎 (TBEV) 和裂谷热 (RVFV) 病毒，以及马中的西尼罗河病毒 (WNV)。所选病毒代表黄病毒属（LIV、TBEV 和 WNV）和白蛉病毒属（RVFV）。这些病毒经过精心挑选，代表了新兴病毒人畜共患病血清监测的复杂性，以证明 SPAr 测定的潜力。与现有的血清监测方法相比，多重SPAr检测具有相当大的优势。它不仅限于单一病原体方法，而且能够区分非常密切相关的病原体，这一特性对于制定人畜共患病原体的有效控制策略至关重要。

项目成果

Phage Display - Methods and Protocols

噬菌体展示 - 方法和方案

• DOI: 10.1007/978-1-0716-3381-6_25
• 发表时间: 2023
• 作者: Tsoumpeli M