Quantitative real-time PCR method for detection of B-lymphocyte monoclonality by comparison of κ and λ immunoglobulin light chain expression
Journal article, 2003

Background: An abnormal IgLκ:IgLλ ratio has long been used as a clinical criterion for non-Hodgkin B-cell lymphomas. As a first step toward a quantitative real-time PCR-based multimarker diagnostic analysis of lymphomas, we have developed a method for determination of IgLκ:IgLλ ratio in clinical samples. Methods: Light-up probe-based real-time PCR was used to quantify IgLκ and IgLλ cDNA from 32 clinical samples. The samples were also investigated by routine immunohistochemical analysis and flow cytometry. Results: Of 32 suspected non-Hodgkin lymphoma samples analyzed, 28 were correctly assigned from real-time PCR measurements assuming invariant PCR efficiencies in the biological samples. Four samples were false negatives. One was a T-cell lymphoma, one was a diffuse large B-cell lymphoma, and one was reanalyzed and found lymphoma-positive by in situ calibration, which takes into account sample-specific PCR inhibition. Twelve of the samples were fine-needle aspirates, and these were all correctly assigned. Conclusions: This work is a first step toward analyzing clinical samples by quantitative light-up probe-based real-time PCR. Quantitative real-time PCR appears suitable for high-throughput testing of cancers by measuring expression of tumor markers in fine-needle aspirates.


Anders Ståhlberg

Chalmers, Department of Chemistry and Bioscience, Molecular Biotechnology

Pierre Åman

University of Gothenburg

Börje Ridell

University of Gothenburg

Petter Mostad

University of Gothenburg

Chalmers, Department of Mathematical Statistics

Mikael Kubista

Chalmers, Department of Chemistry and Bioscience, Molecular Biotechnology

Clinical Chemistry

Vol. 49 1 51-59

Subject Categories

Biochemistry and Molecular Biology

Areas of Advance

Life Science Engineering (2010-2018)

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