Rapid diagnostic biosensors for the detection of respiratory viruses (VIRASENS)

呼吸道合胞病毒（RSV）和其他病毒，如流感，是广泛和严重的感染，特别是在非常年轻和非常年老的人中。例如，RSV在其早期阶段与细菌感染非常相似，但所需的治疗是根本不同的，例如抗病毒或抗细菌。无论哪种方式，错误的决定都会导致时间损失和并发症，这些都可以通过早期准确诊断来避免。因此，这些疾病是需要床旁（POC）诊断的主要例子，而不是需要将样本送到中心实验室的系统。POC有一系列方法，包括简单的侧流装置，如妊娠试验中使用的装置。Orla Protein Technologies Ltd是一家利用BBSRC资助的研究成果设计敏感表面的公司，该表面允许电子设备检测生物分子。这些表面被用于各种系统，但在这项研究中，他们采用了表面声波传感器与日本领先的无线通信公司，日本无线电公司合作制造。该设备，表面声波（或SAW）传感器是一个电子版本的众所周知的音叉与自己定义的音符或谐振频率。如果小分子结合到设备的表面，那么这种特征音符就会改变，并且可以被电子检测到。然后，信号与患者信息、时间、温度甚至GPS阅读一起无线发送到中央设备沿着。因此，结果被自动记录，避免了人为错误。医生可以立即得到答案，并能快速、适当地开出处方。所有这一切都取决于检测的质量，该检测依赖于5 nm厚的层，该层可识别病毒的特征蛋白。5纳米比一张标准A4纸薄30，000倍，因此该层是分子尺度工程的一个例子。Orla使用分子自组装成一个定向层，由于他们的设计，因此在设备上的组装是以一种比油漆/想象油漆一个房间更容易的方式完成的，只需在墙上扔一桶丝绸乳液，并最终得到一个完美的层，避免木制品，天花板和地板！关键在于混合物的分子设计，其中大部分是蛋白质工程。蛋白质是我们希望识别的目标，通常我们使用其他蛋白质来识别它们。其中最好的例子是在小鼠中产生的抗体，它们被选择用于识别RSV或其他病毒蛋白。该小组的研究项目将创建工程蛋白质，这些蛋白质将与抗体共同组装，以在SAW设备上创建高度选择性的单层。这涉及修改自组装蛋白质，以使非常稳定和精确定向的抗体结合阵列，通过生物物理方法检查这些蛋白质的结构，在模型表面上组装蛋白质，如SAW器件上存在的蛋白质，并测量层的所得行为。该公司将在潜在客户手中的真实的生活情况下对设备进行测量。因此，Lakey小组将专注于该层的分子工程，并将通过获得蛋白质工程方面的设施和专业知识为该项目增加价值。作为初步步骤，我们还将使用我们的生物化学知识从维乐通提供的无菌材料中纯化病毒抗原。这些对于质量控制目的是必不可少的，并将为校准和提高设备的灵敏度和选择性提供重要的资源。该项目的最终结果将是一个高度选择性和敏感的自组装层，可以直接转移到商业SAW器件的大规模制造。

Respiratory syncytial virus (RSV) and others, such as influenza, are widespread and serious infections especially important in the very young and the very old. RSV for example presents in its early stages much like bacterial infections but the treatment required is fundamentally different e.g. anti /viral or anti bacterial. The wrong decision, either way, leads to lost time and complications which can be avoided by early accurate diagnosis. These diseases are thus prime examples of the need for point of care (POC) diagnosis rather than systems which require samples to be sent to a central laboratory. There are a range of approaches to POC including simple lateral flow devices like those used in pregnancy tests. Orla Protein Technologies Ltd is a company using the results from BBSRC funded research to design sensitive surfaces which allow electronic devices to detect biological molecules. These surfaces are used in a variety of systems but for this study they are employing a surface acoustic wave sensor manufactured in collaboration with a leading Japanese wireless communication firm, the Japan Radio Company. The device, a surface acoustic wave (or SAW) sensor is an electronic version of the well known tuning fork with its own defined note or resonant frequency. If small molecules bind to the surface of the device then this characteristic note changes and can be detected electronically. The signal is then sent wirelessly to a central device along with patient information, time, temperature and even a GPS reading. Thus the results are automatically logged and human error is avoided. The physician gets an immediate answer and can prescribe quickly and appropriately.All this depends upon the quality of the detection which relies upon a 5 nm thick layer which recognises characteristic proteins from the virus. Five nanometres is 30,000 times thinner than a sheet of standard A4 paper and the layer is thus an example of molecular scale engineering. Orla uses molecules which self-assemble into an oriented layer due to their design, thus the assembly upon the device are accomplished in a manner easier than painting / imagine painting a room by simply throwing a bucket of silk emulsion on the walls and ending up with a perfect layer that avoids the woodwork, ceiling and floor! The trick to this is in the molecular design of the mixture and much of this is protein engineering. Proteins are the target that we wish to recognise and usually we use other proteins to recognise them. The best examples of these are antibodies, produced in mice, which have been selected to recognise RSV or other viral proteins. The research project in this group will create engineered proteins that will co assemble with the antibodies to create a highly selective monolayer on the SAW device. This involves modifying the self assembling proteins to make very stable and precisely oriented antibody binding arrays, checking the structure of these proteins by biophysical methods, assembly of the proteins on model surfaces like the ones present on the SAW device and measurement of the resulting behaviour of the layers. The company will carry out measurements upon the devices under real life situations in the hands of prospective customers. The Lakey group will thus concentrate on the molecular engineering of the layer and will add value to the project by its access to facilities and expertise in protein engineering. As a preliminary step we will also use our biochemical knowledge to purify viral antigens from sterile materials supplied by VIRATOM. These are essential for quality control purposes and will provide an important resource to calibrate and improve the sensitivity and selectivity of the device. The final result of the project will be a highly selective and sensitive self assembling layer which can be transferred directly into the mass manufacture of the commercial SAW device.