A homogenous bimodal (immuno/PCR) pathogen detection system based on a bio-nanoparticle

基于生物纳米颗粒的同质双峰（免疫/PCR）病原体检测系统

• 批准号: BB/J02001X/1
• 负责人: Timothy Dafforn
• 金额: $ 15.28万
• 依托单位: University of Birmingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FJ02001X%2F1
• 关键词: homogenous; bimodal; immuno; PCR; pathogen

Since ancient times armies have attempted to employ biological weapons in order to defeat a foe. Early examples involved tainting water supplies with dead animals or catapulting plague ridden corpses over the walls of besieged cities. In more recent times biological and chemical weapons have become the focus of rogue states and terrorist organisations. Perhaps most high profile of these was the anthrax attacks that occurred in the US in 2001 killing 5 people. Since the UK renounced the use of biological weapons in 1975 it has increased efforts to improve detection of such biowarfare agents. In this project we aim to develop a simple rapid system that will detect biological weapons more quickly and effectively, improving the defence and security of the UK.The system that we will develop is an addition to an existing pathogen detection system that we have pioneered in Birmingham. The existing system relies on viruses to detect the pathogen of choice. These viruses have a spaghetti like structure which means they align with respect to one another when solutions containing them are stirred or forced through narrow tubes. We have modified these viruses so that they are also able to adhere specifically to pathogens; when bound to pathogens the viruses are no longer able to align. The modification of these viruses and their use in an assay is a core part of a new area of science called synthetic biology. This new discipline aims to use biological materials to enhance the performance of everyday devices. In this case we are using synthetic biology to enhance biowarfare detection.The assay involves detecting the changes in the alignment of the viruses and translating this information into a signal that can be read by a soldier or security operative. This area of synthetic biology poses a significant challenge as we need to translate a signal from the virus (which is less 1,000,000th of a metre long) into a signal that can be read. To achieve this we use a simple, yet underused, method (linear dichroism spectroscopy) that is able to detect whether the viruses are aligned or not. If they are aligned then there is no pathogen present, if they are not aligned then the pathogen is present. The method for carrying out the assay has a number of useful characteristics that makes it ideally suited for use by the security services. The assay is simple and very quick allowing the it to be carried out without the need for laboratory equipment. The assay can also be configured so that more than one pathogen can be detected in a single assay. This is important as there are a wide range of biowarfare agents that need to be detected at the same time. Unfortunately, however our current method is only able to use one of the available pathogen detection modes, based on using antibodies, in order to carry out detection.In this project we aim to address this insufficiency by adding a second detection mode to our system. This mode involves directly detecting and identify the DNA from the biowarfare agent. To add this method to our system we have to add small pieces of DNA that match those in biowarfare agent to the surface of the virus. These DNA fragments specifically stick to the DNA in the biowarfare agent preventing the virus from aligning and hence giving us a signal. If this is successful we also aim to add a second step that uses an enzyme to amplify the signal (in a method called PCR) which will make the whole system much more sensitive. In the final stage of the project we will combine the DNA based assay with the antibody based assay to produce a system that is more effective, more flexible and less prone to false results than any current system. In this way we will also demonstrate one of the first uses of synthetic biology in a detection application.

自古以来，军队就试图使用生物武器来击败敌人。早期的例子包括用动物尸体污染水源，或者把瘟疫缠身的尸体扔到被围困的城市的城墙上。近年来，生物和化学武器已成为流氓国家和恐怖组织的焦点。也许其中最引人注目的是2001年在美国发生的炭疽袭击，造成5人死亡。自1975年联合王国宣布放弃使用生物武器以来，它已加紧努力，改进对这种生物战剂的探测。在这个项目中，我们的目标是开发一个简单的快速系统，将检测生物武器更快，更有效，提高英国的国防和安全。我们将开发的系统是一个现有的病原体检测系统，我们已经率先在伯明翰。现有的系统依赖于病毒来检测选择的病原体。这些病毒具有意大利面条状结构，这意味着当含有它们的溶液被搅拌或被迫通过狭窄的管道时，它们会彼此对齐。我们已经对这些病毒进行了改造，使它们也能够特异性地粘附在病原体上;当与病原体结合时，病毒不再能够对齐。对这些病毒的修饰及其在检测中的应用是一个新的科学领域的核心部分，称为合成生物学。这门新学科旨在使用生物材料来提高日常设备的性能。在这种情况下，我们使用合成生物学来增强生物战检测，该检测包括检测病毒排列的变化，并将此信息转化为士兵或安全人员可以读取的信号。合成生物学的这一领域提出了一个重大挑战，因为我们需要将来自病毒（不到1，000，000米长）的信号转化为可以读取的信号。为了实现这一目标，我们使用了一种简单但未充分使用的方法（线性二色性光谱法），该方法能够检测病毒是否对齐。如果它们对齐，则不存在病原体，如果它们不对齐，则存在病原体。用于进行分析的方法具有许多有用的特征，使得其理想地适合于由安全服务使用。该测定方法简单且非常快速，允许在不需要实验室设备的情况下进行。该测定还可以被配置为使得可以在单个测定中检测多于一种病原体。这一点很重要，因为有多种生物战剂需要同时检测。不幸的是，然而，我们目前的方法只能使用一种可用的病原体检测模式，基于使用抗体，以进行检测。在这个项目中，我们的目标是通过添加第二种检测模式到我们的系统来解决这个不足。该模式涉及直接检测和识别来自生物战剂的DNA。为了将这种方法添加到我们的系统中，我们必须在病毒表面添加与生物战剂中的DNA相匹配的小片段。这些DNA片段特异性地粘附在生物战剂中的DNA上，阻止病毒对齐，从而给我们一个信号。如果这是成功的，我们还打算增加第二步，使用酶来放大信号（在一种称为PCR的方法中），这将使整个系统更加敏感。在该项目的最后阶段，我们将联合收割机将基于DNA的检测与基于抗体的检测结合起来，以产生一个比任何当前系统更有效、更灵活且更不容易出现错误结果的系统。通过这种方式，我们还将展示合成生物学在检测应用中的首批用途之一。