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The highly significant role that acquired genetic abnormalities play in the genesis, diagnosis, and management of hematological malignancies has become increasingly clear. Such abnormalities can serve as useful markers for initial diagnosis, accurate subclassification, and the evaluation of minimal residual disease, as well as providing critical targets for novel therapies. In Myeloid Leukemia: Methods and Protocols, a panel of internationally recognized research scientists and clinical investigators brings together a diverse collection of readily reproducible methods for identifying and quantifying a large number of specific genetic abnormalities associated with the broad spectrum of myeloid malignancies. The methods range from those that are of immediate clinical relevance to the investigation and management of patients with myeloid malignancies, to those that relate to recently identified genetic abnormalities of potential clinical significance. Highlights include techniques for thedetection of BCR-ABL mutations and resistance to imatinib mesylate, detection of the FIP1L1-PDGFRA fusion in idiopathic hypereosinophilic syndrome and chronic eosinophilic leukemia, classification of AML by DNA-oligonucleotide microarrays, and detection of the V617F JAK2 mutation in myeloproliferative disorders. In addition to gene rearrangments, other prognostically relevant molecular lesions such as FLT3 mutations and WT-1 overexpression are covered. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Clinically relevant and highly practical, Myeloid Leukemia: Methods and Protocols offers cytogeneticists, hematologists, and oncologists cutting-edge laboratory techniques that can be rapidly implemented for the investigation and management of patients with myeloid malignancies.
Sommario
"Isolation of RNA and DNA From Leukocytes and cDNA Synthesis
Joop H. Jansen and Bert A. van der Reijden
Cytogenetic and FISH Techniques in Myeloid Malignancies
Lynda J. Campbell
Overview of Real-Time RT-PCR Strategies for Quantification of Gene Rearrangements in the Myeloid Malignancies
Christophe Picard, Monique Silvy, and Jean Gabert
Diagnosis and Monitoring of Chronic Myeloid Leukemia by Qualitative and Quantitative RT-PCR
Susan Branford and Timothy Hughes
Detection of BCR-ABL Mutations and Resistance to Imatinib Mesylate
Susan Branford and Timothy Hughes
Deletion of the Derivative Chromosome 9 in Chronic Myeloid Leukemia
Lynda J. Campbell
Diagnosis and Monitoring of PML-RARA-Positive Acute Promyelocytic Leukemia by Qualitative RT-PCR
Vincenzo Rossi, Laura Levati, and Andrea Biondi
Diagnosis and Monitoring of PML-RARa-Positive Acute Promyelocytic Leukemia by Quantitative RT-PCR
Elisa Mokany, Alison V. Todd, Caroline J. Fuery, and Tanya L. Applegate
Diagnosis and Monitoring of AML1-MTG8 (ETO)-Positive Acute Myeloid Leukemia by Qualitative and Real-Time Quantitative RT-PCR
Khalid Tobal and John A. Liu Yin
Diagnosis and Monitoring of CBFB-MYH11-Positive Acute Myeloid Leukemia by Qualitative and Quantitative RT-PCR
Bert A. van der Reijden and Joop H. Jansen
Detection of the FIP1L1-PDGFRA Fusion in Idiopathic Hypereosinophilic Syndrome and Chronic Eosinophilic Leukemia
Jan Cools, Elizabeth H. Stover, and D. Gary Gilliland
FLT3 Mutations in Acute Myeloid Leukemia
Hitoshi Kiyoi and Tomoki Naoe
WT-1 Overexpression in Acute Myeloid Leukemia and Myelodysplastic Syndromes
Daniela Cilloni, Enrico Gottardi, and Giuseppe Saglio
Classification of AML by DNA-Oligonucleotide Microarrays
Alexander Kohlmann, Wolfgang Kern, Wolfgang H iddemann, and Torsten Haferlach
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Riassunto
A panel of internationally recognized research scientists and clinical investigators brings together a diverse collection of readily reproducible methods for identifying and quantifying a large number of specific genetic abnormalities associated with the broad spectrum of myeloid malignancies. Highlights include techniques for the detection of BCR-ABL mutations and resistance to imatinib mesylate, detection of the FIP1L1-PDGFRA fusion in idiopathic hypereosinophilic syndrome and chronic eosinophilic leukemia, classification of AML by DNA-oligonucleotide microarrays, and detection of the V617F JAK2 mutation in myeloproliferative disorders. In addition to gene rearrangments, other prognostically relevant molecular lesions such as FLT3 mutations and WT-1 overexpression are covered.
