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We optimized the assay using HIV-infected cell lines then applied it to bloodstream examples from 18 individuals living with HIV on antiretroviral treatment, yielding a complete of 1,308 distinct HIV-1 genomes. Benchmarking against the widely applied Full-Length Individual Proviral Sequencing assay revealed comparable sensitivity (11% vs 18%) and overall good concordance, though at a significantly greater throughput. In closing, HIV-PULSE is a cost-efficient and scalable assay that allows for the cha