Efficient detection of leukemia-related fusion transcripts by multiplex PCR applied on a microelectronic platform.

ABSTRACT

The identification of prognostically relevant fusion genes is required in the routine diagnostic process of most advanced clinical protocols for leukemia patients, either for risk stratification, target-specific treatments, and/or as markers for monitoring Minimal Residual Disease during treatment. However, there is emerging need to implement diagnostics and patient classification based on other biological features, such as expression levels of specific genes or genomic polymorphisms and/or mutations. This advancement would ideally be pursued in a diagnostic laboratory by an unique platform capable of different diagnostic purposes. We developed a rapid, accurate and reproducible assay to screen for the most common fusion gene transcripts in human leukemia, which combines a multiplex RT-PCR approach with the electronic hybridization and fluorescent detection on the Nanogen NanoChip Molecular Biology Workstation. This study demonstrates, as a proof-of-principle, that this microelectronic device, highly effective in detecting single base mutations, is also efficient in the analysis of gene expression, thus providing as a multi-purpose platform for relevant comprehensive diagnostics of hemato-oncology patients.