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Quantitative real-time PCR for cancer detection: the lymphoma case.

Review article
Authors Anders Ståhlberg
Neven Zoric
Pierre Åman
Mikael Kubista
Published in Expert review of molecular diagnostics
Volume 5
Issue 2
Pages 221-30
ISSN 1473-7159
Publication year 2005
Published at Institute of Laboratory Medicine, Dept of Pathology
Pages 221-30
Language en
Links dx.doi.org/10.1586/14737159.5.2.221
Keywords Gene Expression Profiling, methods, Gene Expression Regulation, Leukemic, Humans, Lymphoma, diagnosis, genetics, Reverse Transcriptase Polymerase Chain Reaction, methods, Tumor Markers, Biological
Subject categories Medical and Health Sciences

Abstract

Advances in the biologic sciences and technology are providing molecular targets for diagnosis and treatment of cancer. Lymphoma is a group of cancers with diverse clinical courses. Gene profiling opens new possibilities to classify the disease into subtypes and guide a differentiated treatment. Real-time PCR is characterized by high sensitivity, excellent precision and large dynamic range, and has become the method of choice for quantitative gene expression measurements. For accurate gene expression profiling by real-time PCR, several parameters must be considered and carefully validated. These include the use of reference genes and compensation for PCR inhibition in data normalization. Quantification by real-time PCR may be performed as either absolute measurements using an external standard, or as relative measurements, comparing the expression of a reporter gene with that of a presumed constantly expressed reference gene. Sometimes it is possible to compare expression of reporter genes only, which improves the accuracy of prediction. The amount of biologic material required for real-time PCR analysis is much lower than that required for analysis by traditional methods due to the very high sensitivity of PCR. Fine-needle aspirates and even single cells contain enough material for accurate real-time PCR analysis.

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