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and 
Immunoglobulin Light Chain Expression
Departments of
1
Molecular Biotechnology and
3
Mathematics, Chalmers University of Technology, 405 30 Gothenburg, Sweden.
2 Department of Pathology, Lundberg Laboratory for Cancer Research, Gothenburg University, 413 45 Gothenburg, Sweden.
4 TATAA Biocenter, Chalmers University of Technology, 405 30 Gothenburg, Sweden.
aAddress correspondence to this author at: TATAA Biocenter, Medicinarg. 9C, Chalmers University of Technology, 405 30 Gothenburg, Sweden. Fax 46-31-7733948; e-mail mikael.kubista{at}tataa.com.
Background: An abnormal IgL
:IgL
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 IgL:IgL ratio in clinical samples.
Methods: Light-up probe-based real-time PCR was used to quantify IgL and IgL 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.
The following articles in journals at HighWire Press have cited this article:
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