Design of standardized "real-time" PCR (RQ-PCR) assays, suitable for "LightCycler" and ABI platforms for genes already known to be commonly over-expressed in leukemias and novel fusion gene targets;
Validation of RQ-PCR assays;· Development of a standardized protocol for MRD detection of RNA-based targets;
Evaluation of computer software programs for RQ-PCR data analysis and reporting;
Interaction with Cytogenetics (WP), Gene Profiling (WP) and Diagnostic platform (WP) to identify potential novel targets for MRD detection
Description of work
Design of standardized RQ-PCR assays, suitable for "Lightcycler" and ABI platforms for genes commonly over-expressed in leukemias and novel fusion gene targets optimization: Comparison of Network designed assays and "in house" sets for sensitivity and reproducibility, using cell line RNA and plasmid standards Design and validation of pre-analytical methods for RNA stabilization and extraction to be used in multicenter trials.
Validation of RQ-PCR assays: Testing in primary leukemia samples, normal bone marrow, peripheral blood leukocytes, regenerating marrow.
Development of a standardized protocol for MRD detection of RNA-based targets: Considering methods for bedside RNA stabilization, standardized reverse transcription and PCR steps.
Evaluation of computer software programs for RQ-PCR data analysis and reporting: Evaluation in on-going clinical trials, including clinician feedback to optimize reporting methods.
The objective of the European LeukemiaNet is to integrate the leading leukemia trial groups (CML, AML, ALL, CLL, MDS, CMPD), their interdisciplinary partners (diagnostics, treatment research, registry, guidelines), industry and SMEs across Europe to form a cooperative network for advancements in leukemia-related research and health care and cure.