BME-IBSC PhD Defense Announcement for Emeka Nwanochie (Jacqueline Linnes and Tamara Kinzer-Ursem, Co-Chairs) Everyone is invited to attend the public presentation beginning at 10:00 AM. Title: TOWARDS QUANTITATIVE MOLECULAR ISOTHERMAL AMPLIFICATION FOR POINT-OF-CARE HIV VIRAL LOAD MONITORING Date: April 12, 2024 Time: 10 AM Location: Purdue Graduate Student Center (PGSC) Room 105 A and B Zoom Link: https://purdue-edu.zoom.us/my/jlinnes<https://nam04.safelinks.protection.outlook.com/?url=https%3A%2F%2Fpurdue-edu.zoom.us%2Fmy%2Fjlinnes&data=05%7C02%7Cbmegradstudents-list%40ecn.purdue.edu%7C449a5f5a268b40072d6a08dc502f05f6%7C4130bd397c53419cb1e58758d6d63f21%7C0%7C0%7C638473410087546686%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C0%7C%7C%7C&sdata=PGg4z9rsv96ddSWm9c7FaBk%2FPoLsyKfWu9npZl7VnEM%3D&reserved=0> (open to all) Committee: 1. Dr. Jacqueline Linnes (Co-Chair) Weldon School of Biomedical Engineering 1. Dr. Tamara Kinzer-Ursem (Co-Chair) Weldon School of Biomedical Engineering 1. Dr. J. Paul Robinson (Member) Weldon School of Biomedical Engineering 1. Dr. Aman Russom (Member) Department of Nanobiotechnology KTH Royal Institute of Technology, Stockholm, Sweden Abstract: The quantification of viral load in people with HIV (PLHIV) is crucial for assessing the effectiveness of antiretroviral therapy and evaluating transmission risk. However, in 2022, 11.3 million PLHIV had still not achieved viral suppression and may become susceptible to both HIV transmission and a variety of opportunistic infections. Nucleic acid amplification tests (NAATs) have emerged as potent tools for monitoring viral load with reverse transcription quantitative polymerase chain reaction (RT-qPCR) being recognized as the benchmark due to its sensitivity and ability for real-time quantification enabled by fluorescence signal emission. Nevertheless, RT-qPCR is burdened by drawbacks including extended processing times, high operational costs, and the requirement for specialized laboratory facilities. In this study, we propose a novel method for HIV-1 viral load monitoring by integrating reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) with real-time particle diffusometry (PD). By monitoring changes in diffusivity during RT-LAMP amplification of HIV-1, real-time PD allows for the generation of quantitative data embedded within PD plots. Additionally, to address challenges related to amplification inhibition in complex human specimens, we developed a power-free sample processing system specifically designed for extracting HIV-1 RNA from both whole blood and plasma. Ultimately, we incorporated the real-time quantitative PD-RT-LAMP assay onto a field-compatible handheld portable platform suitable for field use, featuring built-in quality control measures. These innovations aim to facilitate quick and comprehensive viral load determination, offering promise for enhanced HIV management and patient care.