PROJECT SUMMARY/ABSTRACT The World Health Organization (WHO) recommends algorithms to implement ART regimens, to monitor efficacy by viral load (VL) testing, and for overall HIV management of people living with HIV (PLH). In many locations, this means infrequent VL testing (e.g., annually), and elevated VL typically results in counseling and retesting after several months. Persistent high VL may qualify PLH for drug resistance (DR) testing to differentiate between adherence problems and HIVDR (e.g., in South Africa, eligible for DR testing only after 2 years of failure). This approach can lead to long periods when viremic PLH can spread HIV that could also have DR. DTG-based therapies are a breakthrough in treatment, with low cost and high barrier to resistance, however real-world studies have begun to find emerging DTG resistance. Infrequent VL testing, and delayed or inaccessible DR testing, greatly increase the opportunity to spread DR HIV, which could undermine the effectiveness and longevity of DTG-based therapies. To address this gap, WHO is drafting a Target Product Profile (TPP) that calls for a test that can “determine the need to switch ART (i.e., distinguish between lack of DR suggesting non- adherence, and presence of DR as the reason for treatment failure)”. Such a test should prevent unnecessary switching of patients with treatment failure due to adherence and prevent prolonged viremia in people with DR. To address this need we plan to develop the following: Aim 1: Rapid, low-cost HIV RNA isolation from a large volume of whole blood. Aim 2: Rapid, low-cost HIV viral load test. Aim 3: Rapid, low-cost reflex point-mutation test for HIV drug resistance. Aim 4: Simplified library preparation for efficient semi-centralized nanopore sequencing.