Hiv Rna Isolation
Hiv Rna Isolation - Related Videos
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.
06:46
Isolation of Exosomes from the Plasma of HIV-1 Positive Individuals
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Cited by 51 •
2016
Techniques describing a gradient procedure to separate exosomes from human immunodeficiency virus (HIV) particles are described. This procedure was used to isolate exosomes away from HIV particles in human plasma from HIV-infected individuals. The isolated exosomes were analyzed for cytokine/chemokine content.
05:46
Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors
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Cited by 15 •
2014
Here we describe cellular cytotoxicity and single round infectivity assays that allow for the rapid and accurate screening of compounds to determine their cellular cytotoxicity (CC50) and IC50 values against WT and drug resistant HIV-1.
13:47
Development of Cell-type specific anti-HIV gp120 aptamers for siRNA delivery
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Cited by 36 •
2011
Several 2’-Fluoro RNA aptamers against HIV-1Ba-L gp120 with nanomole affinity are isolated from a RNA library by in vitro SELEX procedure. A new dual inhibitory function anti-gp120 aptamer-siRNA chimera is created and shows considerable promise for systemic anti-HIV therapy.
09:43
Isolation and Characterization of RNA-Containing Exosomes
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Cited by 515 •
2012
This paper demonstrates methods for the isolation, purification and detection of exosomes, as well as techniques for analysis of their molecular content. These methods are adaptable for exosome isolation from both cell culture media and biological fluids, and can beyond analysis of molecular content also be useful in functional studies.
FAQs
Chemical lysis performs two essential functions: it disrupts HIV viral particles and inactivates RNases, enzymes that can degrade RNA. This creates a lysate suitable for the subsequent capture step. Effective lysis and RNase control help preserve the genetic material needed for reverse transcription and amplification.
Following lysis, viral RNA selectively binds to either a silica membrane or magnetic beads. Washing removes unwanted components while the RNA remains associated with the capture surface. An elution step then releases the purified material into a form suitable for downstream molecular analysis, including reverse transcription and amplification.
RNase control helps prevent degradation of the viral RNA during processing. Maintaining recoverable RNA is important because the isolated material must support later reverse transcription and amplification. If the genetic material is not adequately preserved, subsequent measurements or characterization may not reliably reflect the HIV RNA present in the original clinical specimen.
A typical workflow begins with chemical lysis of the clinical specimen, which disrupts viral particles and inactivates RNases. The released RNA is then selectively bound to a silica membrane or magnetic beads, washed to remove unwanted material, and eluted. The recovered RNA proceeds to downstream molecular procedures such as reverse transcription and amplification.
Purified HIV RNA can support viral-load measurement, molecular diagnostics, genotyping, and treatment monitoring. It also provides material for examining viral diversity. Together, these applications allow investigators and clinicians to measure or characterize infection and evaluate changes associated with therapeutic response.
Within immunology and infection research, dependable RNA recovery supplies molecular evidence about HIV infection dynamics and viral variation. Researchers can use the material for genotyping and related characterization, while treatment-monitoring applications help assess therapeutic response. The isolation step therefore connects clinical specimens with measurements of infection status and viral behavior.