Spectrophotometric approaches to develop a bacteriophage based assay to identify pathogens of poultry

分光光度法开发基于噬菌体的检测方法来识别家禽病原体

• 批准号: 1645594
• 金额: --
• 依托单位: University of Leicester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1645594
• 关键词: Spectrophotometric; approaches; develop; bacteriophage; based

Chickens have been shown to carry a wide range of bacterial pathogens that can infect humans. The most important and prevalent of these pathogens are Clostridium perfringens, Salmonella spp, and Campylobacter jejuni,. Detecting what bacterial species the chickens carry is not straightforward and most tests rely on culture, ELISA or molecular approaches which all have problems with specificity and selectivity. Delay in identifying the pathogens, or misidentifying them can lead to the chickens receiving ineffective antibiotics, this has implications for bacterial carriage, animal welfare and the development of antibiotic resistance. The aim of this project is to develop a prototype phage based diagnostic tool that could be used by vets/farmers in order to establish the identity of bacterial pathogens that infect chickens in a quick and effective manner.This project will exploit the specificity of interaction between bacteria and their specific bacteriophages. We will aim to identify phages for each of the major chicken pathogens, and label them, or their tail fibre proteins, with a unique detectable marker. This will be in conjunction with a specifically developed microfluidic device able to produce droplets. Biological samples will then be tested for the pathogens of interest will be passed through the device. If two phage reporters that infect one pathogen attach to a particular cell within a single droplet, there will be a specific co-localised signal unique to that pathogen. This signal will be detected and the pathogen identified with the use of spectroscopy. The development of this assay would have a significant impact on bacterial diagnosis for many situations, however the immediate aim is to address a current and real problem affecting the safety of the food chain, this research is a clear example of strong science which leads to a translational endpoint.

鸡已被证明携带多种可以感染人类的​​细菌病原体。这些病原体中最重要和最普遍的是产气荚膜梭菌、沙门氏菌和空肠弯曲菌。检测鸡携带的细菌种类并不简单，大多数测试依赖于培养、ELISA 或分子方法，这些方法都存在特异性和选择性问题。延迟识别病原体或错误识别病原体可能会导致鸡接受无效的抗生素，这对细菌携带、动物福利和抗生素耐药性的发展产生影响。该项目的目的是开发一种基于噬菌体的原型诊断工具，可供兽医/农民使用，以便快速有效地确定感染鸡的细菌病原体的身份。该项目将利用细菌与其特定噬菌体之间相互作用的特异性。我们的目标是识别每种主要鸡病原体的噬菌体，并用独特的可检测标记来标记它们或其尾部纤维蛋白。这将与专门开发的能够产生液滴的微流体装置结合使用。然后，生物样本将通过该设备进行目标病原体测试。如果感染一种病原体的两个噬菌体报告基因附着在单个液滴内的特定细胞上，就会出现该病原体特有的特定共定位信号。将使用光谱学检测该信号并识别病原体。该测定的开发将对许多情况下的细菌诊断产生重大影响，但直接目标是解决影响食物链安全的当前实际问题，这项研究是导致转化终点的强大科学的明显例子。