Abstract
Human immunodeficiency virus-1 (HIV-1) persistence in the presence of antiretroviral therapy (ART) has halted the development of curative strategies. Measuring HIV persistence is complex due to the low frequency of cells containing virus in vivo. Most of the commercially available assays to date measure nucleic acid. These assays have the advantage of being highly sensitive and allow for the analysis of sequence diversity, intactness of the HIV genome or evaluation of diverse RNA species. However, these assays are limited in evaluating translational competent viral reservoirs. In here, we developed an ultrasensitive p24 ELISA that uses the Simoa planar array technology that can detect HIV-1 virions and HIV-1 infected cells with limit of detection similar to nucleic acid assays. Furthermore, the assay is optimized to measure very low levels of p24 in different biological fluids without a major loss of sensitivity or reproducibility. Our results demonstrate that the ‘homebrew’ planar p24 ELISA immunoassay is a broadly applicable new tool to evaluate HIV persistence in diverse biological fluids and cells.
Highlights
Human immunodeficiency virus-1 (HIV-1) persistence is responsible for the increase in viremia observed after antiretroviral (ART) interruption and can be a driver of residual immune activation observed in (ART)-treated people living with HIV (PLHW)[1]
To further investigate the limit of detection of HIV-1 RNA copies, we evaluated a range of concentrations spanning 1 to 70 copies/ml
The development of ultrasensitive assays to measure HIV-1 protein in diverse biological fluids can add to the repertoire of assays used to evaluate the efficacy of different clinical interventions towards an HIV-1 cure
Summary
Human immunodeficiency virus-1 (HIV-1) persistence is responsible for the increase in viremia observed after antiretroviral (ART) interruption and can be a driver of residual immune activation observed in (ART)-treated people living with HIV (PLHW)[1]. These assays are really effective to evaluate HIV-1 persistence and allow for the analysis of integrated virus, sequence diversity, intactness of the HIV genome or evaluation of diverse RNA species, including splicing forms or poly-A m RNA9–13 These assays are limited in evaluating translational competent viral reservoirs. We have evaluated a homebrew p24 immunoassay developed using the ultrasensitive Quanterix Simoa planar array technology and the SP-X imaging and analysis system[23] The advantages of this new platform are that: (1) it can be performed using volumes between 25 to 50 μl in a 96-well format; (2) it can be optimized and developed to the protein of interest; (3) it allows for the direct quantification in different matrixes without further manipulation; and (4) the full assay can be performed in less than 5 hour. The homebrew planar SP-X immunoassay p24 ELISA can be an additional assay to evaluate HIV-1 cure approaches when sample volume is limited, and to study HIV-1 persistence in diverse anatomical compartments
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
