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Background: Historically, soil-transmitted helminth (STH) control and prevention strategies have relied on mass drug administration efforts targeting preschool and school-aged children. While these efforts have succeeded in reducing morbidity associated with STH infection, recent modeling efforts have suggested that expanding intervention to treatment of the entire community could achieve transmission interruption in some settings. Testing the feasibility of such an approach requires large-scale clinical trials, such as the DeWorm3 cluster randomized trial. In addition, accurate interpretation of trial outcomes requires diagnostic platforms capable of accurately determining infection prevalence (particularly as infection intensity is reduced) at large population scale and with significant throughput. Here, we describe the development and validation of such a high-throughput molecular testing platform., Methodology/principal Findings: Through the development, selection, and validation of appropriate controls, we have successfully created and evaluated the performance of a testing platform capable of the semi-automated, high-throughput detection of four species of STH in human stool samples. Comparison of this platform with singleplex reference assays for the detection of these same pathogens has demonstrated comparable performance metrics, with index assay accuracy measuring at or above 99.5% and 98.1% for each target species at the level of the technical replicate and individual extraction respectively. Through the implementation of a rigorous validation program, we have developed a diagnostic platform capable of providing the necessary throughput and performance required to meet the needs of the DeWorm3 cluster randomized trial and other large-scale operational research efforts for STH., Conclusions/significance: Resulting from the rigorous developmental approach taken, the platform we describe here provides the needed confidence in testing outcomes when utilized in conjunction with large-scale efforts such as the DeWorm3 trial. Additionally, the successful development of an evaluation and validation strategy provides a template for the creation of similar diagnostic platforms for other neglected tropical diseases., Competing Interests: Co-author Sitara Swarna Rao Ajjampur currently sits on the editorial board for PLoS NTDs., (Copyright: © 2025 Pilotte et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)