Rapid Enzymatic Assay for Antiretroviral Drug Monitoring Using CRISPR-Cas12a-Enabled Readout

• DOI: 10.1021/acssynbio.4c00674
• Publication Date: 2025-02-11T20:07:46Z
• Authors: Maya A. Singh; Megan M. Chang; Qin Wang; Catherine Rodgers; Barry R. Lutz; Ayokunle O. Olanr...
• Keywords: Antiretroviral drug

Maintaining the efficacy of human immunodeficiency virus (HIV) medications is challenging among children because dosing difficulties, limited number approved drugs, and low rates medication adherence. Drug level feedback (DLF) can support dose optimization timely interventions to prevent treatment failure, but current tests are heavily instrumented centralized. We developed REverse transcriptase ACTivity crispR (REACTR) for rapid measurement HIV drugs based on extent DNA synthesis by reverse transcriptase. CRISPR-Cas enzymes bind synthesized DNA, triggering collateral cleavage quenched reporters generating fluorescence. measured azidothymidine triphosphate (AZT-TP), a key drug in pediatric treatment, investigated impact assay time template length REACTR's sensitivity. REACTR selectively clinically relevant AZT-TP concentrations presence genomic peripheral blood mononuclear cell lysate. has potential enable point-of-care DLF improve care.