Molecular methods used for detection of minimal residual disease following hematopoietic stem cell transplantation in myeloid disorders.
Monitoring of minimal residual disease (MRD) in patients with acute or chronic myeloid disorders is routinely performed 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 reverse-transcription (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 detection of disease-specific genetic mutated alterations as FLT3 gene-length mutations, and aberrantly expressed genes as WT1 gene transcripts, are described in detail for daily use.
Department of Bone Marrow Transplantation, University Hospital of Essen, Essen, Germany.
Ahmet H
AH
Elmaagacli
D002880:Chromosomes, Human, Pair 11; D002888:Chromosomes, Human, Pair 19; D002896:Chromosomes, Human, Pair 6; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics; D016162:Genes, Wilms Tumor; D016313:Genes, abl; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007951:Leukemia, Myeloid / Q000175:diagnosis / Q000473:pathology / Q000628:therapy*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D018365:Neoplasm, Residual / Q000175:diagnosis*; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D000075142:RUNX1 Translocation Partner 1 Protein; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014178:Translocation, Genetic; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics
C107844:AML1-ETO fusion protein, human; C095514:CBFbeta-MYH11 fusion protein; D050676:Core Binding Factor Alpha 2 Subunit; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3