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

BGR开发了一种直接从粗样品中检测病毒病原体的方法，无需复杂的实验室设施或专家使用。该技术最初是在2015年埃博拉疫情爆发后开发的，以确定是否有可能在资源贫乏的环境中进行实验室标准分子诊断。这方面的建议是调整联邦遗传资源研究所的技术，使其从检测病毒性出血热到检测艾滋病毒等病毒载量较低但影响人数多得多的疾病。在该项目中，BGR将建造一个新的反应容器，用于进行管内检测，从而能够使用更大体积的血液，从而提高灵敏度。BGR开发的试剂能够在高水平血液存在下检测病毒RNA，迄今为止高达20%，但在这些高百分比下，用于确定目标存在的光学数据受到抑制。这项工作的目标是创建后期原型血管，结合进化的试剂，将血液从光学光路中去除，从而提高方法的灵敏度，并大大增加该方法适用的疾病数量。例如，埃博拉患者每毫升血液中可能有一百万个病毒，而丙型肝炎患者每毫升血液中可能有五万个病毒，而艾滋病患者每毫升血液中可能有一万个病毒。目标是将灵敏度提高到3000/ml，世卫组织已确定5000/ml适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方法只需不到50分钟，不需要实验室设施或培训，成本低，因为每个测试都由塑料管和冻干酶组成。因此，改进的反应容器、试剂和仪器将大大增加这项创新对健康的影响，并为公司带来商业机会。然后，该测定可用于协助血液传播疾病的爆发情况。还设想为该项目开发的新容器可以使用其他类型的样本，例如用于呼吸道疾病的拭子。然后，BGR可以使用模型病毒确定新检测方法的灵敏度，并将适当的检测方法商业化。

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.