Real-Time Visualization of HIV-1 RNA Detection Using Loop-Mediated Isothermal Amplification-Enabled Particle Diffusometry.

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  • Additional Information
    • Source:
      Publisher: American Chemical Society Country of Publication: United States NLM ID: 101669031 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2379-3694 (Electronic) Linking ISSN: 23793694 NLM ISO Abbreviation: ACS Sens Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, DC : American Chemical Society, [2016]-
    • Subject Terms:
      HIV-1*/genetics ; HIV-1*/isolation & purification ; RNA, Viral*/analysis ; Nucleic Acid Amplification Techniques*/methods; Diffusion ; Humans ; Molecular Diagnostic Techniques
    • Abstract:
      Isothermal nucleic acid amplification tests, NAATs, such as reverse transcription-loop-mediated isothermal amplification (RT-LAMP), offer promising capabilities to perform real-time semiquantitative detection of viral pathogens. These tests provide rapid results, utilize simple instrumentation for single-temperature reactions, support efficient user workflows, and are suitable for field use. Herein, we present a novel and robust method for real-time monitoring of HIV-1 RNA RT-LAMP utilizing a novel implementation of particle diffusometry (PD), a diffusivity quantification technique using fluorescent particles, to quantify viral concentration in nuclease-free water. We monitor changes in particle diffusion dynamics of 400 nm fluorescently labeled particles throughout the RT-LAMP of HIV-1 RNA in nuclease-free water, enabling measurement within 20 min and detection of concentrations as low as 25 virus particles per μL. Moreover, in a single-blind study, we demonstrate semiquantitative detection by accurately determining the initial concentration of an unknown HIV-1 RNA within a 10% absolute error margin. These results highlight the potential of real-time PD readout for quantifying HIV-1 RNA via RT-LAMP, offering promise for viral load monitoring of HIV and other chronic infections.
    • Contributed Indexing:
      Keywords: RT-LAMP; diagnostics; diffusion coefficient; fluorescence; isothermal amplification; particle diffusometry; real time
    • Accession Number:
      0 (RNA, Viral)
    • Subject Terms:
      LAMP assay
    • Publication Date:
      Date Created: 20241008 Date Completed: 20241025 Latest Revision: 20241025
    • Publication Date:
      20241026
    • Accession Number:
      10.1021/acssensors.4c01870
    • Accession Number:
      39377655