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

Anders Ståhlberg (Institutionen för kemi och biovetenskap, Molekylär bioteknik) ; Pierre Åman ; Börje Ridell ; Petter Mostad (Institutionen för matematisk statistik) ; Mikael Kubista (Institutionen för kemi och biovetenskap, Molekylär bioteknik)
Clinical Chemistry Vol. 49 (2003), 1, p. 51-59.
[Artikel, refereegranskad vetenskaplig]

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.



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Denna post skapades 2012-07-02. Senast ändrad 2015-08-25.
CPL Pubid: 159900

 

Institutioner (Chalmers)

Institutionen för kemi och biovetenskap, Molekylär bioteknik (2002-2004)
Institutionen för laboratoriemedicin , Avdelningen för patologi (1991-2005)
Institutionen för matematisk statistik (2002-2004)

Ämnesområden

Livsvetenskaper
Molekylärbiologi

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