RAPID: Highly Specific and Sensitive Detection of Ebola Virus from Blood

1509759 -Santangelo为了减轻埃博拉病毒感染的传播，在症状出现之前检测患者体内的病毒势在必行。 这就需要对病毒进行快速、灵敏和特异的检测。 目前的方法缺乏灵敏度和易用性。 该项目将采用作者以前的方法检测患者血液中超低浓度的埃博拉病毒。 将开发两种新方法;一种可用于现场，第二种更严格，适用于实验室测试。这些方法有望降低埃博拉疫情的传播速度。目前，定量PCR是检测埃博拉病毒的最先进方法，通常从血液样本中进行检测。 然而，这些测试是时间密集型的，需要高度的专业知识来执行。 此外，当患者体内的病毒浓度相对较高时，该方法是有用的。鉴于作为隔离策略的一部分，需要对大量人群进行检测，因此改进早期诊断势在必行。因此，该项目的目的是开发一种更灵敏的方法来检测埃博拉病毒RNA（vRNA）和单个病毒颗粒。研究小组将开发一种适合现场应用的单管检测全血中病毒RNA的方法。此外，还将通过检测vRNA-蛋白质相互作用，开发用于检测单个病毒颗粒的基于玻片的检测方法，该方法具有适用于实验室环境的单个病毒粒子灵敏度。在这两种情况下，将使用肽标记的单个RNA敏感的RNA成像探针和邻位连接测定（PLA）方法。 将结果与当前方法进行比较，以证明灵敏度的提高。 位点特异性探针结合和距离依赖性检测和扩增的组合将确保特异性（零误差）和灵敏度（10拷贝/ml）。

1509759 - SantangeloIn order to mitigate the spread of Ebola infections, detecting the virus in patients before the onset of symptoms is imperative. This requires, fast, sensitive and specific detection of the virus. Current methods lack sensitivity and ease of use. This project will adapt the author's previous method for detecting ultra low Ebola concentration in patient's blood. Two new methods will be developed; one can be used in the field and the second one, which is more rigorous, is suitable for laboratory tests. These methods are expected to reduce the rate at which Ebola epidemic spreads.Currently quantitative PCR is the state-of-the-art for Ebola detection, typically from blood samples. These tests though, are time intensive and require a high degree of expertise to perform. Further the method is useful when viral concentration is relatively high in the patient. Given the need to test large numbers of people as a part of quarantine strategies, improved early diagnostics are imperative. Therefore the aim of the project is to develop a more sensitive approach to detecting Ebola viral RNA (vRNA) and single viral particles. The research team will develop a single tube assay for detecting viral RNA in whole blood, suitable for field application. In addition, a slide-based assay for detecting single viral particles will also be developed, through the detection of vRNA-protein interactions, with single virion sensitivity suitable for a laboratory setting. In both cases, peptide labeled, single RNA sensitive, RNA imaging probes and proximity ligation assay (PLA) methods will be utilized. The results will be compared with current methods to demonstrate the improved sensitivity. The combinations of both site-specific probe binding and distance-dependent detection and amplification, will ensure both specificity (zero errors) and sensitivity (10 copies/ml).