Assessment of minimal residual disease in B-cell chronic lymphocytic leukemia: options and advancement of techniques based on PCR and RT-PCR


Authors: H. Skuhrová Francová;  B. Tichý;  K. Malinová;  J. Mayer;  Š. Pospíšilová
Authors‘ workplace: Centrum molekulární biologie a genové terapie, Interní hematoonkologická klinika Lékařské fakulty MU a FN Brno
Published in: Transfuze Hematol. dnes,15, 2009, No. 4, p. 197-203.
Category: Comprehensive Reports, Original Papers, Case Reports

Overview

Detection of minimal residual disease (MRD) persistence provides essential information on the treatment response in hematologic malignancies including chronic lymphocytic leukemia (CLL). Improved quantitative techniques for MRD quantification on molecular basis with sufficient sensitivity are warranted to monitor the therapeutical goal - complete erradication of MRD. In addition to international standardized four-color flow cytometric CLL-MRD assay, real time polymerase chain reaction (RQ-PCR) represents one of the most sensitive methods for MRD assesment. The malignant clone can be identified by its unique immunoglobulin heavy chain (IgH) gene rearrangement; allele-specific (ASO) IgH RQ-PCR allows to achieve higher sensitivity limit up to two orders of magnitude. Currently, the monitoring of MRD is based on combination of patient specific primers and two different consensus TaqMan probes recognizing the JH subgenes 1, 4, 5, 6 and is applicable to just about 90 % of all CLL cases. The updated approach is based on limited number of LNA-modified (Locked Nucleic Acid) TaqMan probes designed to consensus sequences in the framework region 3 (FR3) of the seven VH gene families. LNA-modified probes and IgH-RQ PCR represent highly specific and sensitive method of MRD assessment in patiens with CLL.

Key words:
chronic lymphocytic leukemia (CLL), immunoglobulin heavy-chain gene (IgVH), minimal residual disease (MRD), real-time quantitative PCR (RQ PCR)


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Haematology Internal medicine Clinical oncology
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