Validity test study of JAK2 V617F and allele burden quantification in the diagnosis of myeloproliferative diseases.
Several sensitive methods for the detection of JAK2 V617F mutation have been published recently, most of them based on Real Time polymerase chain reaction (PCR). However, only some of them have performed studies of diagnostic validity. This study compares three methods based on Real Time PCR to detect JAK2 V617F mutation: two based on hybridization probes (HP) and peptide nucleic acid probe (PNA) and a third employing allele specific oligonucleotide primers for JAK2 V617F quantification. One hundred forty-nine healthy subjects, 61 essential thrombocythemia (ET), 32 polycythemia vera (PV), 38 secondary thrombocytoses, and 35 secondary erythrocytoses were included. Validity test study for JAK2 617 HP PCR in PV Sensitivity (Se) was 88% and in Specificity (Sp), 100%. In ET, Se was 57% and Sp, 100%. For JAK2 617 PNA PCR in PV, Se was 94% and Sp, 97.8%. In ET, Se was 70% and Sp, 95.7%. In JAK2 V671F allelo-specific-oligonucleotide (ASO) quantitative PCR (qPCR), cutoff point of 1% was established by receiving operating characteristic (ROC) curves. In PV, Se was 93.8% and Sp, 98.5%. In ET, Se was 80% and Sp, 95.9%. Two percent of the healthy subjects were positive by JAK2 617 PNA PCR and 2% by JAK2 617 ASO qPCR. JAK2 V617F mutation was detected in healthy subjects by cloning and sequencing. JAK2 617 HP is an adequate test in differential diagnosis for both erythrocytosis and thrombocytosis. When JAK2 V617F allele burden is low, JAK2 617 ASO qPCR should be performed. Simultaneous determination of JAK2 V617F and PRV-1 overexpression does not improve the diagnostic value of JAK2 V617F tests in MPD.
Hematology Service, Hospital Universitario 12 de Octubre, Avenida Córdoba s/n, 28041 Madrid, Spain.
Inmaculada
I
Rapado
Enriqueta
E
Albizua
Rosa
R
Ayala
Jose Angel
JA
Hernández
Luis
L
Garcia-Alonso
Silvia
S
Grande
Miguel
M
Gallardo
Florinda
F
Gilsanz
Joaquin
J
Martinez-Lopez
D019943:Amino Acid Substitution; D017931:DNA Primers; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000097:blood / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D011086:Polycythemia / Q000235:genetics; D016133:Polymerase Chain Reaction; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013816:Thermodynamics; D013922:Thrombocytosis / Q000235:genetics
D017931:DNA Primers; C507924:JAK2 protein, human; D053614:Janus Kinase 2