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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Related Experiment Video

Updated: Jun 3, 2026

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
13:07

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells

Published on: January 30, 2019

Quantitation of Cell-Free HIV by Reverse Transcriptase Activity.

J R Lane1

  • 1SRA Technologies, Inc., Rockville, MD.

Methods in Molecular Medicine
|March 8, 2011
PubMed
Summary
This summary is machine-generated.

This assay detects reverse transcriptase (RT) activity, an indicator of HIV-1 presence in human cell cultures. Measuring RT in cell supernatants helps confirm active viral infection in vitro.

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Last Updated: Jun 3, 2026

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
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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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Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays

Published on: September 26, 2011

Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Reverse transcriptase (RT) is a key enzyme in retroviral replication, synthesizing DNA from an RNA template.
  • Human immunodeficiency virus type 1 (HIV-1) infection is characterized by the activity of its own RT.
  • Active HIV-1 infection leads to the release of virions and associated enzymes into the cell culture medium.

Purpose of the Study:

  • To describe an assay for measuring reverse transcriptase activity.
  • To establish RT activity as a marker for HIV-1 presence in vitro.
  • To outline the method for isolating and concentrating viral particles from cell culture supernatants for analysis.

Main Methods:

  • The assay quantifies the enzymatic activity of reverse transcriptase (RNA-dependent-DNA polymerase).
  • Samples are derived from supernatants of human peripheral blood mononuclear cells (PBMCs) cultured in vitro.
  • Virus is concentrated using polyethylene glycol (PEG) precipitation and resuspended for assay.

Main Results:

  • Detection of RT activity in cell culture supernatants indicates the presence of HIV-1.
  • The assay allows for the identification of active viral replication in vitro.
  • Concentration and purification steps enhance the sensitivity of RT detection.

Conclusions:

  • The reverse transcriptase assay is a reliable method for detecting HIV-1 in vitro.
  • RT activity serves as a direct indicator of active viral infection in PBMC cultures.
  • The described methodology facilitates the sensitive detection of HIV-1 through RT enzyme activity.