Molecular methods used for detection of minimal residual disease following hematopoietic stem cell transplantation in myeloid disorders.

The monitoring of minimal residual disease (MRD) in patients with acute or chronic myeloid disorders is performed routinely after allogeneic or autologous transplantation. The detection of MRD helps to identify patients who are at high risk for leukemic relapse after transplantation. The most commonly used techniques for MRD detection are qualitative and quantitative PCR methods, fluorescence in situ hybridization (FISH), fluorescence-activated cell sorting (FACS), and cytogenetic analysis, which are often performed complementary in order to assess more precisely MRD. Here we describe the most used sensitive real-time RT-PCR methods for chronic and acute myeloid disorders. Besides protocols for real-time RT-PCR and multiplex RT-PCR procedures for the most common fusion-gene transcripts in acute and chronic myeloid disorders, methods for detections of disease-specific genetic mutated alterations, as NPM1 and FLT3 gene length mutations, and aberrantly expressed genes, as WT1 gene transcripts, are described in detail for daily use.