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Title: An overview of real-time quantitative PCR: applications to quantify cytokine gene expression
Authors: Giulietti, Annapaula *
Overbergh, Lutgart *
Valckx, Dirk
Decallonne, Brigitte
Bouillon, Roger
Mathieu, Chantal # ×
Issue Date: Dec-2001
Publisher: Academic Press
Series Title: Methods vol:25 issue:4 pages:386-401
Abstract: The analysis of cytokine profiles helps to clarify functional properties of immune cells, both for research and for clinical diagnosis. The real-time reverse transcription polymerase chain reaction (RT-PCR) is becoming widely used to quantify cytokines from cells, body fluids, tissues, or tissue biopsies. Being a very powerful and sensitive method it can be used to quantify mRNA expression levels of cytokines, which are often very low in the tissues under investigation. The method allows for the direct detection of PCR product during the exponential phase of the reaction, combining amplification and detection in one single step. In this review we discuss the principle of real-time RT-PCR, the different methodologies and chemistries available, the assets, and some of the pitfalls. With the TaqMan chemistry and the 7700 Sequence Detection System (Applied Biosystems), validation for a large panel of murine and human cytokines and other factors playing a role in the immune system is discussed in detail. In summary, the real-time RT-PCR technique is very accurate and sensitive, allows a high throughput, and can be performed on very small samples; therefore it is the method of choice for quantification of cytokine profiles in immune cells or inflamed tissues.
URI: 
ISSN: 1046-2023
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Clinical and Experimental Endocrinology
Research Group Experimental Neurology
* (joint) first author
× corresponding author
# (joint) last author

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