Simultaneous screening for JAK2 and calreticulin gene mutations in myeloproliferative neoplasms with high resolution melting.
BACKGROUND
Recently, novel calreticulin (CALR) mutations were discovered in Janus kinase 2 (JAK2) non-mutated myelofibrosis (PMF) and essential thrombocythemia (ET) cases, with a frequency of 60-80%. We examined clinical correlations and CALR mutation frequency in our myeloproliferative neoplasms (MPN) cases, and introduce an effective test method for use in clinical practice.


METHODS
We examined 177 samples previously investigated for the JAK2 mutation for differential diagnosis of MPN. JAK2 and CALR mutations were analyzed using melting curve analysis and microchip electrophoresis, respectively. Next, we constructed a test for simultaneous screening of the JAK2 and CALR mutations utilizing high resolution melting (HRM).


RESULTS
Among 99 MPN cases, 60 possessed the JAK2 mutation alone. Of the 39 MPN cases without the JAK2 mutation, 14 were positive for the CALR mutation, all of which were ET. Using our novel screening test for the JAK2 and CALR mutations by HRM, the concordance rate of conventional analysis with HRM was 96% for the JAK2 mutation and 95% for the CALR mutation.


CONCLUSIONS
Our novel simultaneous screening test for the JAK2 and CALR gene mutations with HRM is useful for diagnosis of MPN.
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan.
Nariyoshi
N
Matsumoto
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan.
Sayaka
S
Mori
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan; Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan. Electronic address: hhase@nagasaki-u.ac.jp.
Hiroo
H
Hasegawa
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan.
Daisuke
D
Sasaki
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan.
Hayato
H
Mori
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan.
Kazuto
K
Tsuruda
Nagasaki Goto Chuo Hospital, Nagasaki, Japan.
Daisuke
D
Imanishi
Department of Hematology, Nagasaki University Hospital, Nagasaki, Japan.
Yoshitaka
Y
Imaizumi
Department of Hematology, Nagasaki University Hospital, Nagasaki, Japan; Atomic Bomb Disease and Hibakusha Medicine Unit, Atomic Bomb Disease Institute, Nagasaki University, Japan.
Tomoko
T
Hata
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan; Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Norihito
N
Kaku
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan; Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Kousuke
K
Kosai
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan; Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Naoki
N
Uno
Department of Hematology, Nagasaki University Hospital, Nagasaki, Japan; Atomic Bomb Disease and Hibakusha Medicine Unit, Atomic Bomb Disease Institute, Nagasaki University, Japan.
Yasushi
Y
Miyazaki
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan; Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Katsunori
K
Yanagihara
D037282:Calreticulin / Q000235:genetics*; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000175:diagnosis / Q000235:genetics; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000175:diagnosis / Q000235:genetics*
C576483:CALR protein, human; D037282:Calreticulin; C507924:JAK2 protein, human; D053614:Janus Kinase 2
Calreticulin; High resolution melting; Janus kinase 2 V617F; Myeloproliferative neoplasms; Screening test