Objective
Objectives:
1. Optimization and standardization of PCR primer sets and PCR techniques for detection of Ig/TcR gene rearrangements and well-defined chromosome aberrations as targets for clonality assessment.
2. Evaluation of the applicability of optimized and standardized PCR techniques for clonality studies in all NHL categories as defined by the Revised European American Lymphoma (REAL) classification.
Brief description:
The molecular diagnosis of NHL is based on the fact that all cells of a malignancy have a common clonal origin. This is reflected by the presence of identically (clonally) rearranged immunoglobulin (Ig) and T-cell receptor (TcR) genes in >98% of NHL and the presence of well-defined chromosome aberrations in 25-30% of NHL. Clonal Ig/TcR gene rearrangements and most well-defined chromosome aberrations in NHL can be detected by Southern blot analysis and PCR-based techniques. Southern blotting is considered to be the gold-standard technique for clonality studies. However, PCR-based techniques are preferred, because they are rapid and require minimal amounts of medium-quality DNA, which might even be obtained from small formalin-fixed paraffin-embedded tissue samples. Two major drawbacks hamper the application of PCR-based techniques for analysis of Ig/TcR gene rearrangements. Firstly, relatively high frequencies of false-negative results (10-30% of cases) are obtained due to improper primer annealing. The second difficulty is the discrimination between PCR products derived from monoclonal and polyclonal cell populations. PCR analysis of chromosome aberrations might be hampered by the fact that the positions of the chromosome breakpoints differ per patient. Therefore most PCR approaches for clonality studies focus on well-defined cluster regions of breakpoints such as mbr and mcr in t(14;18) and MTC in t(11;14). Nevertheless, multiple primer sets are needed for reliable detection of all breakpoints in one cluster region.
Several diagnostic/research groups in Europe have tried to solve the problems of PCR-based clonality studies, but so far no reliably standardized PCR protocols have been obtained. In contrast, many different primer sets are used, which all differ in their sensitivity and applicability. For rapid and reliable diagnosis of lymphoproliferative disorders it is of utmost importance to solve the problems of PCR-based clonality assessment in a European collaborative study by experienced laboratories.
To facilitate the organization and management of this complex and extensive collaboration, it was decided to structure the Concerted Action via seven national networks, each coordinated by a core laboratory. This structure of seven core laboratories and their national networks will stimulate the commitment of each participant, facilitates the monitoring of the progress of the collaborative studies, and enables efficient exchange of technology both at the national level and at the European level.
keywords
Lymphoproliferation, non-Hodgkin's lymphoma (NHL), early diagnosis, PCR-based-clonality-assessment, immunoglobulin (Ig) and T-cell receptor (TcR) genes, chromosome aberrations, international standardization, primer-design, PCR-product-analysis, diagnostic evaluation.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences genetics DNA
- social sciences sociology social issues social inequalities
- natural sciences biological sciences genetics chromosomes
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Coordinator
3015 GE Rotterdam
Netherlands
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