LCVD: Low-cost Cell-extract Viral Diagnostics

南非的病毒疾病负担南非是世界上艾滋病毒感染率最高的国家之一。对于受感染的个体，药物治疗取决于了解他们的病毒载量（VL），即有多少病毒在循环。去年在南澳大利亚州进行了大约240万次艾滋病毒VL检测，每次检测费用为25至40美元，但那些无法进入主要保健中心的人仍然无法进行检测。人类诺如病毒（HuNoV; “冬季呕吐病毒”）具有高度传染性，90%的非细菌性腹泻病是由它引起的。在撒哈拉以南非洲的研究表明，5岁以下儿童与腹泻相关的死亡中有很大一部分没有报告。因此，在南非，人类免疫缺陷病毒感染很可能是一种被低估的健康威胁。用低成本的护理点诊断测试解决这两个问题将大大改善诊断和治疗，降低死亡率，并减轻这些疾病沉重的经济负担。提案的目的和目标我们的最终目标是共同开发方法，以快速设计和构建具有成本效益的护理点诊断方法，用于导致南非和其他低收入地区高发病率和死亡率的传染病病原体。我们将以HIV病毒载量（VL）检测和人类诺如病毒（HuNoV）诊断为目标。最近，基于生物的寨卡病毒和埃博拉病毒的“生物传感器”测试已经开发出来，可以在纸条上生产。这些测试有可能非常低成本，因为它们使用细胞的汤或提取物来识别病毒。这种“无细胞”提取液可以被标记在纸条上，这样就可以对病毒进行一种“试纸”生物传感器测试。我们计划通过一个由英国和南非研究人员组成的团队共同开发HIV和HuNoV的无细胞生物传感器来完善这一基本过程。共同开发将培训SA研究人员开发方法，而且还允许将该方法传授给其他研究人员，然后这些研究人员可以将该方法应用到他们自己的研究中。因此，我们的目标是为南非和其他低收入地区共同开发一种可持续发展的技术。我们的目标是：1)生产用于HIV VL检测和HIV诊断的低成本护理点诊断测试原型；2)使用共同开发研究过程生成一套无细胞生物传感器方法，作为SA和其他LMIC研究人员和教育工作者可访问的平台技术；3)使用共同开发研究过程广泛培训个人研究人员（我们的研究团队），并教授这些方法并培训SA的其他研究人员。我们的提议是基于共同开发无细胞体外生物传感器诊断，利用我们的研究团队来推动英国和南非实验室和人员的创新和整合。

Virus disease burden in South AfricaSouth Africa has one of the highest rates of HIV infection worldwide. For infected individuals, drug therapy depends on knowing their virus load (VL) that is, how much virus is in circulation. About 2.4 million HIV VL tests costing $25 to $40 each, were performed last year in SA but the test is still unavailable those without access to major health care centres. Human norovirus (HuNoV; "winter vomiting virus") is highly infectious and responsible for 90% of non-bacterial diarrheal disease and studies in sub-Saharan Africa suggest that a significant proportion of diarrhea-related deaths in children under 5 go unreported. Thus it is likely that HuNoV infection is an underestimated health threat in SA. Addressing both of these problems with low cost point-of care diagnostic tests would greatly improve diagnosis and treatment, reducing mortality and also the heavy economic burden these diseases.Aims and goals of the proposalOur ultimate aim is to co-develop methods to rapidly design and construct cost-effective point-of-care diagnostics for infectious disease agents responsible for high morbidity and mortality in South Africa and other low-income regions. We will target HIV virus load (VL) testing and human norovirus (HuNoV) diagnosis. Recently, biologically-based "biosensor" tests for Zika virus and Ebola virus have been developed that can be produced on paper strips. These tests have the potential to be very low cost because they use a soup or extract from cells to identify the virus. The "cell-free" extract can be spotted onto paper strips, which allows a type of "dipstick" biosensor test for the virus. We plan to refine this basic process by having a team of UK and SA researchers co-develop the cell-free biosensors for HIV and HuNoV. The co-development will both train SA researchers in method development but also allow the method to be taught to other researchers that can then apply the method to their own research. Hence, we aim to co-develop a sustainable technology for SA and other low-income areas. Our goals are: 1) to produce prototype low cost point-of-care diagnostic tests for HIV VL testing and HuNoV diagnosis 2) to use the co-development research process to generate a set of cell-free biosensor methods as a platform technology accessible to SA and other LMIC researcher and educators 3) to use the co-development research process to broadly train individual researchers (our research team) and teach the methods and train other researchers in SA. Our proposal is based on co-development of cell-free in vitro biosensor diagnostics using our research teamwork to drive innovation and integration of laboratories and personnel in the UK and South Africa.