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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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  S. A. Bustin, V. Benes, J. A. Garson, J. Hellemans, J. Huggett, M. Kubista, R. Mueller, T. Nolan, M. W. Pfaffl, G. L. Shipley, et al. The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments Clin. Chem., April 1, 2009; 55(4): 611 - 622. [Abstract] [Full Text] [PDF]
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 W. Si, J. Gong, Y. Han, H. Yu, J. Brennan, H. Zhou, and S. Chen Quantification of Cell Proliferation and Alpha-Toxin Gene Expression of Clostridium perfringens in the Development of Necrotic Enteritis in Broiler Chickens Appl. Envir. Microbiol., November 1, 2007; 73(21): 7110 - 7113. [Abstract] [Full Text] [PDF]
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 O. Treeck, G. Pfeiler, D. Mitter, C. Lattrich, G. Piendl, and O. Ortmann Estrogen receptor {beta}1 exerts antitumoral effects on SK-OV-3 ovarian cancer cells J. Endocrinol., June 1, 2007; 193(3): 421 - 433. [Abstract] [Full Text] [PDF]
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 K. L. Montooth, K. T. Siebenthall, and A. G. Clark Membrane lipid physiology and toxin catabolism underlie ethanol and acetic acid tolerance in Drosophila melanogaster J. Exp. Biol., October 1, 2006; 209(19): 3837 - 3850. [Abstract] [Full Text] [PDF]
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 S. Ghosh, A. C. Steere, B. D. Stollar, and B. T. Huber In Situ Diversification of the Antibody Repertoire in Chronic Lyme Arthritis Synovium J. Immunol., March 1, 2005; 174(5): 2860 - 2869. [Abstract] [Full Text] [PDF]
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 R. G. Rutledge Sigmoidal curve-fitting redefines quantitative real-time PCR with the prospective of developing automated high-throughput applications Nucleic Acids Res., December 15, 2004; 32(22): e178 - e178. [Abstract] [Full Text] [PDF]
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A. Stahlberg, M. Kubista, and M. Pfaffl Comparison of Reverse Transcriptases in Gene Expression Analysis Clin. Chem., September 1, 2004; 50(9): 1678 - 1680. [Full Text] [PDF]
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A. Stahlberg, J. Hakansson, X. Xian, H. Semb, and M. Kubista Properties of the Reverse Transcription Reaction in mRNA Quantification Clin. Chem., March 1, 2004; 50(3): 509 - 515. [Abstract] [Full Text] [PDF]
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 T. Pastinen, R. Sladek, S. Gurd, A.'a Sammak, B. Ge, P. Lepage, K. Lavergne, A. Villeneuve, T. Gaudin, H. Brandstrom, et al. A survey of genetic and epigenetic variation affecting human gene expression Physiol Genomics, January 15, 2004; 16(2): 184 - 193. [Abstract] [Full Text] [PDF]
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T. Bar, A. Stahlberg, A. Muszta, and M. Kubista Kinetic Outlier Detection (KOD) in real-time PCR Nucleic Acids Res., September 1, 2003; 31(17): e105 - e105. [Abstract] [Full Text] [PDF]
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G. J. Escobar Effect of the Systemic Inflammatory Response on Biochemical Markers of Neonatal Bacterial Infection: A Fresh Look at Old Confounders Clin. Chem., January 1, 2003; 49(1): 21 - 22. [Full Text] [PDF]
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