Project Summary/Abstract We propose the development of a point-of-care LAMP assay for the detection of active HCV infection as a companion to High Performance Plasma (HPP), a novel blood-derived sample type that is advantageous for diagnostic testing when compared to blood or centrifuged plasma. HPP is similar to plasma but also contains PBMCs, which can be used to augment the total quantifiable viral load in the case of active infection, thereby leading to detection at earlier windows than previously possible. By building on existing work in literature, we will develop a sample-to-answer HCV diagnostic assay that is capable of providing accurate diagnostics that rival (and in some cases exceed) the performance of lab testing equivalents within 30 minutes. Beyond HCV, success in this effort can open the door to a variety of POC whole blood NAAT as the Century tube is virtually compatible with any NAAT protocol. Our Phase 1 effort includes three aims which should validate the essential components of a POC LAMP assay. Aim 1: Develop LAMP primers: We will adapt promising primers published in literature to use as part of an ultrasensitive qualitative test to detect HCV. We will conduct our testing in two different types of blood-derived samples: HPP and plasma. Testing will be conducted in spiked samples. Aim 2: Develop Sample Purification Protocol: We will validate the use of a point-of-care compatible centrifuge- free sample purification protocol to extract viral RNA from spiked plasma and HPP samples. Similar to Aim 1, the work will be conducted in spiked samples. Aim 3: Incorporate primers and purification protocol to test human samples: We will combine the results of Aims 1 and 2 to create the framework of a diagnostic LAMP assay. We will then use this assay to test human whole blood samples collected from HCV+ and healthy donors and compare the performance in HPP and plasma.