Information about Project 12: MRD

Created by: Hellenbrecht (ELIC) , generated 2006/02/14, last changed: 2006/11/30

Objectives

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;
Optimization;
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

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.