Myelofibrosis Diagnostics

Laboratory Tests and Ancillaries

Essential Tests

Essential tests that should be requested include a complete blood count (CBC) with differential and platelet count (in which leukocytosis, leukopenia, thrombocytopenia, or thrombocytosis may be seen), peripheral smear, metabolic panel including LDH and serum uric acid levels, liver function tests (LFTs), renal function tests, serum erythropoietin (EPO), and serum iron tests.  

Human leukocyte antigen (HLA) typing is recommended for patients with myelofibrosis when allogeneic hematopoietic stem cell transplant is being considered.  

Biopsy

Bone Marrow Aspirate with Iron Stain and Biopsy  

Bone marrow aspirate with iron stain and biopsy is necessary for the demonstration of fibrosis which is required for the diagnosis of myelofibrosis. Reticulin stain is used to demonstrate bone marrow fibrosis and trichrome stain is used to distinguish between myelofibrosis grades.  

Molecular and Genetic Analysis

Molecular testing for JAK2, CALR, and MPL mutations is recommended for the diagnosis of primary myelofibrosis. Molecular testing using blood or bone marrow for JAK2 V617F mutation is recommended in the initial workup and if negative, molecular testing for MPL and CALR mutations is recommended. Multigene next-generation sequencing (NGS) may be used to test for JAK2, CALR, and MPL mutations. NGS is recommended for mutational prognostication once diagnosis of MF is confirmed. Other clonal markers include ASXL1, EZH2, RAS, TET2, IDH1, IDH2, SRSF2, and SF3B1 gene mutations.

Myelofibrosis_Diagnostics

FISH is performed on peripheral blood to detect BCR-ABL1 transcripts and rule out chronic myeloid leukemia.  

Other Tests

Coagulation Tests

Coagulation tests are performed in patients who will undergo high-risk surgery or those with elevated platelet counts and/or splenomegaly or unexplained bleeding, to evaluate for acquired von Willebrand syndrome and/or other coagulopathies. These tests include prothrombin time (PT), partial thromboplastin time (PTT), and fibrinogen. Plasma von Willebrand Factor Antigen (VWFA) measurement and Von Willebrand Ristocetin Cofactor (VWF:RCo) activity may be used if initial screening tests (eg PT, PTT) are prolonged and cannot be corrected.