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用于动物病原体即时诊断的核酸检测、新平台及纳米技术

Nucleic-acid testing, new platforms and nanotechnology for point-of-decision diagnosis of animal pathogens.

作者信息

Teles Fernando, Fonseca Luís

机构信息

Laboratory of Mycology/Microbiology Unit and Centre for Malaria and Tropical Diseases (CMDT), Institute of Hygiene and Tropical Medicine (IHMT), Universidade Nova de Lisboa (UNL), Lisboa, Portugal,

出版信息

Methods Mol Biol. 2015;1247:253-83. doi: 10.1007/978-1-4939-2004-4_20.

Abstract

Accurate disease diagnosis in animals is crucial for animal well-being but also for preventing zoonosis transmission to humans. In particular, livestock diseases may constitute severe threats to humans due to the particularly high physical contact and exposure and, also, be the cause of important economic losses, even in non-endemic countries, where they often arise in the form of rapid and devastating epidemics. Rapid diagnostic tests have been used for a long time in field situations, particularly during outbreaks. However, they mostly rely on serological approaches, which may confirm the exposure to a particular pathogen but may be inappropriate for point-of-decision (point-of-care) settings when emergency responses supported on early and accurate diagnosis are required. Moreover, they often exhibit modest sensitivity and hence significantly depend on later result confirmation in central or reference laboratories. The impressive advances observed in recent years in materials sciences and in nanotechnology, as well as in nucleic-acid synthesis and engineering, have led to an outburst of new in-the-bench and prototype tests for nucleic-acid testing towards point-of-care diagnosis of genetic and infectious diseases. Manufacturing, commercial, regulatory, and technical nature issues for field applicability more likely have hindered their wider entrance into veterinary medicine and practice than have fundamental science gaps. This chapter begins by outlining the current situation, requirements, difficulties, and perspectives of point-of-care tests for diagnosing diseases of veterinary interest. Nucleic-acid testing, particularly for the point of care, is addressed subsequently. A range of valuable signal transduction mechanisms commonly employed in proof-of-concept schemes and techniques born on the analytical chemistry laboratories are also described. As the essential core of this chapter, sections dedicated to the principles and applications of microfluidics, lab-on-a-chip, and nanotechnology for the development of point-of-care tests are presented. Microdevices already applied or under development for application in field diagnosis of animal diseases are reviewed.

摘要

准确诊断动物疾病对于动物健康至关重要,同时对于预防人畜共患病传播给人类也至关重要。特别是,家畜疾病可能会对人类构成严重威胁,这是因为人与家畜之间身体接触频繁且暴露程度高,而且即使在非流行国家,家畜疾病也可能导致重大经济损失,这些国家的家畜疾病往往以迅速且具有毁灭性的疫情形式出现。快速诊断测试在野外环境中已经使用了很长时间,尤其是在疫情爆发期间。然而,它们大多依赖血清学方法,这种方法可以确认是否接触过特定病原体,但在需要基于早期准确诊断的应急响应的即时决策(即时护理)环境中可能并不适用。此外,它们的灵敏度往往不高,因此很大程度上依赖于中央或参考实验室的后续结果确认。近年来,材料科学、纳米技术以及核酸合成与工程领域取得了令人瞩目的进展,催生了大量用于核酸检测的新型台式和原型测试,旨在实现对遗传和传染病的即时护理诊断。与基础科学差距相比,制造、商业、监管和技术性质等野外适用性问题更有可能阻碍它们更广泛地进入兽医学和实践领域。本章首先概述了用于诊断兽医关注疾病的即时护理测试的现状、要求、困难和前景。随后讨论了核酸检测,特别是即时护理核酸检测。还描述了一系列在概念验证方案和分析化学实验室产生的技术中常用的有价值的信号转导机制。作为本章的核心内容,介绍了致力于微流体、芯片实验室和纳米技术在即时护理测试开发中的原理和应用的章节。回顾了已经应用或正在开发用于动物疾病现场诊断的微型设备。

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