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. 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.

Isolation of RNA and DNA From Leukocytes and cDNA Synthesis.- Cytogenetic and FISH Techniques in Myeloid Malignancies.- Overview of Real-Time RT-PCR Strategies for Quantification of Gene Rearrangements in the Myeloid Malignancies.- Diagnosis and Monitoring of Chronic Myeloid Leukemia by Qualitative and Quantitative RT-PCR.- Detection of BCR-ABL Mutations and Resistance to Imatinib Mesylate.- Deletion of the Derivative Chromosome 9 in Chronic Myeloid Leukemia.- Diagnosis and Monitoring of PML-RARA-Positive Acute Promyelocytic Leukemia by Qualitative RT-PCR.- Diagnosis and Monitoring of PML-RAR?-Positive Acute Promyelocytic Leukemia by Quantitative RT-PCR.- Diagnosis and Monitoring of AML1-MTG8 (ETO)-Positive Acute Myeloid Leukemia by Qualitative and Real-Time Quantitative RT-PCR.- Diagnosis and Monitoring of CBFB-MYH11-Positive Acute Myeloid Leukemia by Qualitative and Quantitative RT-PCR.- Detection of the FIP1L1-PDGFRA Fusion in Idiopathic Hypereosinophilic Syndrome and Chronic Eosinophilic Leukemia.- FLT3 Mutations in Acute Myeloid Leukemia.- WT-1 Overexpression in Acute Myeloid Leukemia and Myelodysplastic Syndromes.- Classification of AML by DNA-Oligonucleotide Microarrays.- Classification of AML Using a Monoclonal Antibody Microarray.- Methods for the Detection of the JAK2 V617F Mutation in Human Myeloproliferative Disorders.- Overexpression of PRV-1 Gene in Polycythemia Rubra Vera and Essential Thrombocythemia.- Chimerism Analysis Following Nonmyeloablative Stem Cell Transplantation.