Building a prototype PCR reaction vessel for the direct detection of pathogens from blood

构建用于直接检测血液中病原体的原型 PCR 反应容器

• 批准号: 104580
• 金额: $ 16.98万
• 依托单位: B G RESEARCH LIMITED
• 依托单位国家: 英国
• 项目类别: Feasibility Studies
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=104580
• 关键词: Building; prototype; PCR; reaction; vessel

BGR has developed a method for detecting viral pathogens directly from crude samples, removing the requirement for complex lab facilities or expert use. The technique was originally developed in the wake of the 2015 Ebola outbreak to establish if it is possible to perform laboratory standard molecular diagnostics in resource poor environments. The proposal here is to adapt the BGR technology from the detection of viral haemorrhagic fevers to diseases such as HIV that have lower viral loads but impact far higher numbers of individuals. In this project BGR will build a new reaction vessel for performing its in tube detection that enables the use of greater volumes of blood and hence increase the sensitivity. The reagents that BGR have developed are capable of detecting the viral RNA in the presence of high levels of blood, as much as 20% to date, and yet at these high percentages the optical data used to determine the presence of the targets are inhibited. The goal of this work is to create late stage prototype vessels that, in combination with evolved reagents, remove the blood from the optical light path and therefore increase the sensitivity of the approach and greatly increase the number of diseases the approach will be appropriate for. As an example, an Ebola patient may have a million viruses per millilitre of blood whereas a patient with hepatitis C will have fifty thousand and a HIV patient on drugs around ten thousand. The target is to increase the sensitivity to 3000/ml, the WHO has identified 5000/ml as being suited to low-cost diagnostics in resource poor regions for the detection of disease such as Hepatitis and HIV. The BGR approach takes less than fifty minutes, requires no laboratory facilities or training and is low cost simply because each test on consists of a plastic tube and a freeze-dried enzyme. Therefore, the improved reaction vessel, reagents and instrument would greatly increase the health impact of this innovation and the commercial opportunities for the company. The assays can then be used to assist in outbreak situations of blood borne diseases. It is also envisaged that the new vessel developed for this project could make possible the use of other sample types such as swabs for respiratory disease. BGR can then determine how sensitive the new assays are using a model virus and move towards commercialisation of appropriate tests.

BGR开发了一种直接从粗样品中检测病毒病原体的方法，从而消除了对复杂实验室设施或专家使用的要求。这项技术最初是在2015年埃博拉疫情爆发后开发的，目的是确定是否有可能在资源匮乏的环境中进行实验室标准分子诊断。这里的建议是将BGR技术从病毒性出血热的检测应用到艾滋病毒等病毒载量较低但影响个体数量大得多的疾病。在这个项目中，BGR将建造一个新的反应容器，用于进行管内检测，从而可以使用更大容量的血液，从而提高灵敏度。BGR开发的试剂能够在高水平的血液中检测到病毒RNA，到目前为止，检测率高达20%，然而在如此高的百分比下，用于确定目标存在的光学数据被抑制了。这项工作的目标是创建后期原型血管，与进化的试剂结合，将血液从光学光路中移除，从而提高该方法的灵敏度，并大大增加该方法适用的疾病数量。例如，一名埃博拉患者每毫升血液中可能含有100万个病毒，而一名丙型肝炎患者每毫升血液中可能含有5万个病毒，一名正在服药的艾滋病患者每毫升血液中可能含有1万个病毒。目标是将灵敏度提高到3000/ml，世界卫生组织已经确定5000/ml适合用于资源贫乏地区的低成本诊断，用于检测肝炎和艾滋病毒等疾病。BGR方法耗时不到50分钟，不需要实验室设施或培训，而且成本低，因为每次测试都由一根塑料管和一种冻干酶组成。因此，改进后的反应容器、试剂和仪器将大大增加这一创新对健康的影响和公司的商业机会。然后，这些检测可用于在血液传播疾病暴发的情况下提供协助。还设想，为该项目开发的新容器可以使用其他类型的样本，如呼吸道疾病的拭子。然后，BGR可以确定使用模型病毒的新检测方法的敏感性，并朝着适当检测方法的商业化迈进。