Hiv-1 Replication Assay
Hiv-1 Replication Assay - Related Videos
10:12
Measuring Endoplasmic Reticulum Stress and Unfolded Protein Response in HIV-1 Infected T-Cells and Analyzing its Role in HIV-1 Replication
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Cited by 1 •
2024
Here, we describe some established methods to determine endoplasmic reticulum (ER) stress and unfolded protein response (UPR) activation, with particular emphasis on HIV-1 infection. This article also describes a set of protocols to investigate the effect of ER stress/UPR on HIV-1 replication and virion infectivity.
13:58
Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays
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Cited by 10 •
2011
Quantifying levels of HIV-1 RNA in plasma and sequencing single HIV-1 genomes from individuals with viral loads below the limit of detection (50-75 copies/ml) is difficult. Here we describe how to extract and quantify plasma viral RNA using a real time PCR assay that reliably measures HIV-1 RNA down to 0.3 copies/ml and how to amplify viral genomes by single genome sequencing, from samples with very low viral loads.
14:23
A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
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Cited by 12 •
2014
HIV-1 pathogenesis is defined by both viral characteristics and host genetic factors. Here we describe a robust method that allows for reproducible measurements to assess the impact of the gag gene sequence variation on the in vitro replication capacity of the virus.
10:49
In vitro Uncoating of HIV-1 Cores
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Cited by 47 •
2011
Uncoating is an essential step in the early phase of the HIV-1 life cycle and is defined as the disassembly of the capsid shell and the release of the viral ribonucleoprotein complex (vRNP). Here, we demonstrate techniques for isolating intact cores from HIV-1 virions and for quantifying their uncoating in vitro.
12:07
Preparation and Use of HIV-1 Infected Primary CD4+ T-Cells as Target Cells in Natural Killer Cell Cytotoxic Assays
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Cited by 12 •
2011
Cytotoxicity assays to measure natural killer cell lytic responses to HIV-infected cells is limited by the purity of the target cells. We demonstrate here the isolation of a highly purified population of HIV-1 infected primary T-cell blasts by taking advantage of HIV-1 s ability to down-modulate CD4.
FAQs
An assay can be aligned with successive points in infection: reverse transcription, genome integration, and later viral production. Measuring a signal after these stages helps researchers ask whether an intervention or viral change affects an early replication event or the generation of new virus. This stage-aware interpretation connects molecular activity with the observed infection outcome.
Viral RNA, capsid protein p24, infectious particles, and reporter signals do not represent identical measurements. RNA or p24 can indicate viral material associated with replication, whereas infectious-particle measurements address production of virus that can participate in infection. Reporter signals provide another way to track replication after reverse transcription. Comparing these readouts helps match the assay to the biological question.
Drug potency is inferred by examining how strongly treatment changes the measured replication signal under the assay conditions. Because the workflow can monitor events from reverse transcription through viral production, measurements can relate reduced output to the broader replication process, rather than relying only on a qualitative observation of infection. Standardized measurements also enable comparisons among treatments.
Researchers begin with susceptible cells and expose them to HIV-1, then monitor replication using a selected indicator. Depending on the experimental goal, that indicator may be viral RNA, p24, infectious particles, or a reporter signal. The resulting measurement is used to compare infection or virus production across the conditions being studied.
Keeping the assay framework consistent allows investigators to examine whether different viral variants produce different replication measurements in susceptible cells. The same approach can test the contribution of host factors by comparing outcomes when those cellular influences differ. Such comparisons connect variation at the viral or cellular level with measurable changes in infection and virus production.
Neutralizing-antibody studies use replication measurements to determine how antibody exposure relates to the amount of infection or viral production detected in the assay. Signals such as RNA, p24, infectious particles, or reporter output provide quantifiable endpoints for comparing antibody effects. In immunology and infection research, this links an immune response to a measurable consequence in the viral replication process.