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Real Time RT-PCR

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...

PCR

PCR

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Related Experiment Video

Updated: Jul 5, 2026

Generation of Human Alloantigen-specific T Cells from Peripheral Blood

Generation of Human Alloantigen-specific T Cells from Peripheral Blood

Published on: November 21, 2014

Assays for alloreactive responses by PCR.

Patrick Stordeur¹

¹Département d'Immunologie-Hématologie-Transfusion, Hôpital Erasme, Université Libre de Bruxelles, Brussels, Belgium.

Methods in Molecular Biology (Clifton, N.J.)

|May 6, 2008

Summary

A new rapid assay monitors immune tolerance after organ transplantation using minimal blood. This method quantizes interferon-gamma and interleukin-2 mRNA levels, aiding immunosuppression withdrawal without rejection.

Area of Science:

Immunology
Transplantation Medicine
Molecular Biology

Background:

Minimizing immunosuppression is crucial in organ transplantation to prevent side effects.
Inducing graft tolerance, allowing allograft survival without ongoing immunosuppression, is a key goal.
Current methods for monitoring immune status are labor-intensive and require large blood volumes, hindering clinical application.

Purpose of the Study:

To develop a rapid, low-volume blood assay for monitoring immune tolerance post-transplantation.
To facilitate the reduction or withdrawal of immunosuppressive drugs without compromising graft survival.
To assess anti-donor reactivity in patients undergoing transplantation.

Main Methods:

A novel real-time polymerase chain reaction (PCR) method quantifying interferon-gamma and interleukin-2 messenger RNA (mRNA).

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Induction of Alloantigen-specific Anergy in Human Peripheral Blood Mononuclear Cells by Alloantigen Stimulation with Co-stimulatory Signal Blockade

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Published on: March 14, 2011

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Published on: April 19, 2017

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Last Updated: Jul 5, 2026

Generation of Human Alloantigen-specific T Cells from Peripheral Blood

Generation of Human Alloantigen-specific T Cells from Peripheral Blood

Published on: November 21, 2014

Induction of Alloantigen-specific Anergy in Human Peripheral Blood Mononuclear Cells by Alloantigen Stimulation with Co-stimulatory Signal Blockade

Induction of Alloantigen-specific Anergy in Human Peripheral Blood Mononuclear Cells by Alloantigen Stimulation with Co-stimulatory Signal Blockade

Published on: March 14, 2011

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Published on: April 19, 2017

A modified mixed lymphocyte reaction (MLR) using whole blood T lymphocytes or peripheral blood mononuclear cells (PBMCs).

The assay requires only 10 ml of blood and provides results within 48 hours.

Main Results:

The developed real-time PCR method successfully quantified specific mRNA levels.
The modified MLR assay demonstrated effectiveness in monitoring anti-donor reactivity.
The technique was successfully applied to living donor liver graft recipients.

Conclusions:

This rapid MLR assay offers a valuable tool for monitoring immune status in transplantation.
The method's efficiency and low blood requirement support its implementation in routine clinical practice.
It aids in assessing graft tolerance and guiding immunosuppression management, potentially reducing rejection risk.