title: Detection of the activating JAK2 V617F mutation in paraffin-embedded trephine bone marrow biopsies of patients with chronic myeloproliferative diseases.
abstract: The discovery of the activating V617F mutation in the JAK2 tyrosine kinase in a high proportion of patients with Ph- chronic myeloproliferative diseases (CMPD) represents a diagnostic breakthrough for these disorders. Trephine bone marrow biopsy is an essential part of the diagnostic workup of CMPD and represents a valuable archival source of DNA. Therefore, we studied 152 paraffin-embedded trephines with CMPD and related disorders for the presence of the V617F mutation, using both allele-specific polymerase chain reaction (PCR) and nested PCR with subsequent digestion with BsaXI. Only 6 of 152 (4%) samples were not evaluable because of poor DNA quality. The V617F mutation was detected in 27 of 28 (96%) cases of polycythemia vera, 17 of 23 (74%) cases of essential thrombocythemia, 28 of 45 (62%) cases of chronic idiopathic myelofibrosis, six of eight (75%) cases of CMPD unclassified, and two of four (50%) cases of myelodysplastic/myeloproliferative syndrome. Ph+ chronic myelogenous leukemia (four cases), reactive (secondary) erythrocytosis (14 cases), and thrombocytosis (one case) as well as normal controls (19 cases) all lacked the V617F mutation. Based on results of BsaXI digestion and sequencing, 24 of 54 (44%) evaluable V617F+ cases were considered homozygously mutated. Thus, detection of the V617F JAK2 mutation is feasible in paraffin-embedded trephine biopsies and represents a major advance in the diagnostic evaluation of CMPD.
authors.affiliation: Institute of Pathology, Technical University Munich, Ismaninger Str. 22, 81675 Munich, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Horn
authors.forename: Marcus
authors.initials: M
authors.lastname: Kremer
authors.forename: Tobias
authors.initials: T
authors.lastname: Dechow
authors.forename: Walther M
authors.initials: WM
authors.lastname: Pfeifer
authors.forename: Birgit
authors.initials: B
authors.lastname: Geist
authors.forename: Michael
authors.initials: M
authors.lastname: Perker
authors.forename: Justus
authors.initials: J
authors.lastname: Duyster
authors.forename: Leticia
authors.initials: L
authors.lastname: Quintanilla-Martinez
authors.forename: Falko
authors.initials: F
authors.lastname: Fend
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001706:Biopsy / Q000379:methods*; D001853:Bone Marrow / Q000473:pathology*; D002908:Chronic Disease; D004252:DNA Mutational Analysis / Q000379:methods*; D005240:Feasibility Studies; D005260:Female; D006801:Humans; D053614:Janus Kinase 2; D008297:Male; D008875:Middle Aged; D009196:Myeloproliferative Disorders / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D016612:Paraffin Embedding / Q000379:methods*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D015183:Restriction Mapping; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA
chemical_list: D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; C507924:JAK2 protein, human; D053614:Janus Kinase 2