Stanovení minimální zbytkové nemoci u B-buněčné chronické lymfocytární leukemie: možnosti a vývoj metodických přístupů založených na PCR a RQ-PCR

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

SKUHROVÁ FRANCOVÁ Hana TICHÝ Boris PLEVOVÁ Karla MAYER Jiří POSPÍŠILOVÁ Šárka

Year of publication 2009
Type Article in Periodical
Magazine / Source Transfuze a hematologie dnes
MU Faculty or unit

Faculty of Science

Citation
Field Oncology and hematology
Keywords chronic lymphocytic leukemia (CLL); imunoglobulin heavy-chain gene (IgVh); minimal residual disease (MRD); real-time quantitative PCR (RQ-PCR)
Description 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 identifed 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 assesment in patients with CLL.
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