Molecular diagnostics are essential in today's management of patients with hematological malignancies. In the current WHO classification of tumors, diseases are defined by morphologic, immunologic and genetic features reflecting the role of molecular diagnostics. This book summarizes the current approaches and impact of molecular diagnostics in the most common hematological malignancies (multiple myeloma, lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, myeloproliferative disorders, chronic myeloid leukemia, and myelodysplastic syndromes). Furthermore, molecular diagnostics do not only provide the basis for disease classification, prognostic assessment and treatment decisions, but also greatly contribute to continuous improvements in our understanding of underlying pathogenic events. Therefore, this book not only summarizes today's standard approaches of molecular diagnostics but also illustrates novel experimental strategies and techniques currently used in research laboratories. This comprehensive state-of-the-art overview was made possible by the invaluable contributions of a distinguished group of international investigators with outstanding expertise and track record in their field.

Contenu
1.Basic Principles of Molecular Diagnostics (S. Pfister, P. Lichter)201.1.Introduction201.2.Polymerase Chain Reaction-based methods201.2.1.Quantitative Real-time PCR (QRT-PCR)201.2.1.1.QRT-PCR using a hydrolysis probe ("TaqMan probe")201.2.1.2.QRT-PCR using a hybridization probe201.2.1.3.QRT-PCR using SYBR green dye201.2.1.4.QRT-PCR Applications211.2.2.PCR-based methods to detect promoter methylation of single genes211.2.2.1.Bisulfite genomic sequencing (BGS)211.2.2.2.Methylation-specific PCR (MSP)211.2.2.3.Combined bisulfite restriction analysis (COBRA)221.2.3.Multiplex Ligation-dependent probe amplification (MLPA)221.3.Cytogenetics221.3.1.Conventional Cytogenetics221.3.2.Fluorescence In Situ hybridization (FISH)221.3.3.Comparative Genomic Hybridization (CGH)231.4.Microarray-based Methods231.4.1.Array-based CGH231.4.2.SNP Profiling231.4.3.Messenger-RNA Expression Profiling241.4.4.Epigenetic profiling241.4.5.Micro-RNA Profiling251.5.Immunostaining and Flow Cytometry251.5.1.Immunocytochemistry251.5.2.Flow Cytometry251.6.Proteomics251.6.1.2D-PAGE251.6.2.Mass spectrometry261.7.Sequencing Methods261.7.1.Pyrosequencing261.7.2.Next-generation sequencing (NGS) approaches261.8.Outlook272.Multiple Myeloma (C. Langer, P. Liebisch, R. Fonseca)302.1.Introduction302.2.Cell Sorting302.3.Chromosome Banding302.4.Molecular Diagnostics302.4.1.High Resolution genome-wide Detection Methods312.4.2.FISH322.4.3.Metaphase spectral karyotype Imaging (SKY)332.4.4.Gene Expression Profiling332.5.Pathogenetic Relevance of chromosomal Abnormalities332.5.1.B-cell Development and Myelomagenesis332.5.2.Primary and secondary IgH Translocations332.5.3.Aneuploidy352.5.4.Genetic Model of Myelomagenesis352.5.5.Implication of GEP Data " TC Classification362.6.Clinical and prognostic Implications of chromosomal Abnormalities362.6.1.Aneuploidy362.6.2.Specific genomic Imbalances362.6.2.1.Monosomy 13/13q Deletion (-13/13q-)362.6.2.2.Chromosome 17p Deletion372.6.2.3.Chromosome 1q Abnormalities372.6.2.4.t(11;14)(q13;q32)372.6.2.5.t(4;14)(p16.3;q32)382.6.2.6.t(14;16)(q32;q23)383.Non-Hodgkin-Lymphoma and Hodgkin Lymphoma (J.I Martín-Subero, A. Rosenwald, R. Siebert, R.D. Gascoyne)423.1.Introduction423.2.Chromosomal Changes associated with non-Hodgkin and Hodgkin Lymphomas423.2.1.B-cell non-Hodgkin Lymphomas433.2.1.1.Follicular Lymphoma433.2.1.2.Mantle Cell Lymphoma433.2.1.3.Diffuse large Cell Lymphoma443.2.1.4.Burkitt Lymphoma443.2.2.T-cell non-Hodgkin Lymphomas453.2.3.Hodgkin Lymphoma453.3.Technical Approaches to detect chromosomal Changes in Lymphomas463.3.1.Conventional Cytogenetics463.3.2.Molecular Genetics463.3.3.Molecular Cytogenetics473.3.3.1.Fluorescence In Situ Hybridization473.3.3.2.Combined interphase Cytogenetics and Fluorescence Immunophenotyping (FICTION technique)473.3.3.3.Array-based Techniques to detect chromosomal Changes483.3.4.Technical Comparison of different Methods for the Detection of chromosomal Changes in Lymphomas483.4.Gene Expression Profiling in Lymphomas493.5.Epigenomic Profiling in Lymphomas513.6.Future Directions524.Chronic Lymphocytic Leukemia (S. Stilgenbauer, T. Zenz, D. Oscier)584.1.Introduction584.2.Chromosome banding584.3.Fluorescence In Situ Hybridization (FISH)584.4.Sequence and Mutation Analysis604.5.Array-based Comparative Genomic Hybridization (Array CGH)614.6.SNP arrays624.7.Next Generation Sequencing (NGS)624.8.Gene expression (mRNA) profiling634.9.MicroRNA profiling634.10.IGHV mutation status by sequencing and surrogate markers for the IGHV status by flow cytometry634.11.Conclusion645.Acute Lymphoblastic Leukemia (M. Brüggemann, M. Kneba, T. Raff, T. Szczepañski, J.J.M van Dongen)685.1.Introduction685.2.Immunologic and molecular diagnostics at diagnosis685.2.1.Immunophenotyping685.2.2.Cytogenetics and molecular Genetics705.2.3.Pharmacogenetics725.2.4.Gene Expression Arrays745.3.Molecular Diagnostics during Treatment " Detection of minimal residual Disease745.3.1.MRD Detection by multiparameter Flow Cytometry745.3.2.MRD Quantification using clonal Immunoglobulin and T-Cell-Receptor Gene Rearrangements755.3.3.MRD Quantification using Fusion Genes as RQ-PCR Targets765.3.4.Methodological Aspects of MRD Monitoring in ALL775.3.5.Clinical Significance of MRD in ALL775.4.Conclusion796.Acute Myeloid Leukemia (L. Bullinger, K. Döhner, J.R. Pollack)846.1.Introduction846.2.Immunophenotyping846.3.Chromosome Banding Analysis (conventional Karyotyping)856.4.Fluoresence In Situ Hybridization (FISH)866.5.Polymerase Chain Reaction (PCR) based Techniques876.6.Molecular Genetics for Mutation Detection/Screening886.7.Altered Gene Expression in AML-gene Expression Profiling (GEP)906.8.Genomics Technologies (Microarray based Technologies)926.8.1.Genome-wide analyses of CNAs and UPDs926.8.2.Altered DNA methylation936.8.3.Histone modifications and leukemogenesis936.9.Conclusion957.The diagnosis of BCR-ABL1 negative myeloproliferative neoplasms (MPNs)(U. Bacher, C. Haferlach, T. Haferlach, A. Hochhaus, W. Kern, S. Schnittger)1027....