Rapid identification of heterozygous or homozygous JAK2 V617F mutations in myeloproliferative neoplasms using melting curve analysis

Ching Liang Ho, Yi Ying Wu, Hsiu Man Hung, Ping Ying Chang, Wei You Kao, Yeu Chin Chen, Tsu Yi Chao

Research output: Contribution to journalArticle

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Abstract

Background/Purpose: The activating JAK2 mutation with a G-C to T-A transversion at codon 617 (JAK2 V617F) is associated with myeloproliferative neoplasms (MPNs), including polycythemia vera (PV), essential thrombocythemia (ET), and myelofibrosis. Here, we report a technical advance in the diagnosis of JAK2 V617F in MPNs by melting curve analysis (MCA). Methods: From January through December 2006, we prospectively enrolled 78 patients with PV (n=21), ET (n=32), myelofibrosis (n=5), secondary erythrocytosis (n=4), secondary thrombocytosis (n=2), acute myelocytic leukemia (n=4), chronic myelocytic leukemia (n=8), and myelodysplastic syndrome (n=2). Mutation analysis for JAK2 V617F was performed on either bone marrow or peripheral blood cells using allele-specific polymerase chain reaction (AS-PCR) or sequencing and fluorescence resonance energy transfer (FRET) probes with MCA. Results: For the initial 30 samples, the detection rate of JAK2 V617F using MCA was comparable to the gold standard of the PCR sequencing methods. However, the turnaround times for MCA and PCR were 2 hours and 2 days, respectively. The detection rate of JAK2 V617F was 76.2% for PV (homozygous in 14.3%), 46.9% for ET, 80% for myelofibrosis (homozygous in 20%), and 0% for the other conditions. In PV, patients with homozygous JAK2 V617F presented with significantly longer disease durations than heterozygous patients. In ET, there were no differences in the clinical parameters of patients harboring JAK2 V617F compared with those with wild-type JAK2. Conclusion: Heterozygous and homozygous JAK2 V617F mutations can be identified using the rapid and reliable assay based on FRET probes and MCA. Detection of JAK2 V617F can be used to assist in the diagnosis of BCR/ABL-negative MPNs.

Original languageEnglish
Pages (from-to)34-40
Number of pages7
JournalJournal of the Formosan Medical Association = Taiwan yi zhi
Volume111
Issue number1
DOIs
Publication statusPublished - Jan 2012

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Essential Thrombocythemia
Polycythemia Vera
Freezing
Primary Myelofibrosis
Mutation
Fluorescence Resonance Energy Transfer
Neoplasms
Polymerase Chain Reaction
Thrombocytosis
Polycythemia
Myelodysplastic Syndromes
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Acute Myeloid Leukemia
Codon
Blood Cells
Bone Marrow
Alleles

Keywords

  • Essential thrombocythemia
  • JAK2
  • Melting curve analysis
  • Myeloproliferative neoplasms
  • Polycythemia vera

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Rapid identification of heterozygous or homozygous JAK2 V617F mutations in myeloproliferative neoplasms using melting curve analysis. / Ho, Ching Liang; Wu, Yi Ying; Hung, Hsiu Man; Chang, Ping Ying; Kao, Wei You; Chen, Yeu Chin; Chao, Tsu Yi.

In: Journal of the Formosan Medical Association = Taiwan yi zhi, Vol. 111, No. 1, 01.2012, p. 34-40.

Research output: Contribution to journalArticle

Ho, Ching Liang ; Wu, Yi Ying ; Hung, Hsiu Man ; Chang, Ping Ying ; Kao, Wei You ; Chen, Yeu Chin ; Chao, Tsu Yi. / Rapid identification of heterozygous or homozygous JAK2 V617F mutations in myeloproliferative neoplasms using melting curve analysis. In: Journal of the Formosan Medical Association = Taiwan yi zhi. 2012 ; Vol. 111, No. 1. pp. 34-40.
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abstract = "Background/Purpose: The activating JAK2 mutation with a G-C to T-A transversion at codon 617 (JAK2 V617F) is associated with myeloproliferative neoplasms (MPNs), including polycythemia vera (PV), essential thrombocythemia (ET), and myelofibrosis. Here, we report a technical advance in the diagnosis of JAK2 V617F in MPNs by melting curve analysis (MCA). Methods: From January through December 2006, we prospectively enrolled 78 patients with PV (n=21), ET (n=32), myelofibrosis (n=5), secondary erythrocytosis (n=4), secondary thrombocytosis (n=2), acute myelocytic leukemia (n=4), chronic myelocytic leukemia (n=8), and myelodysplastic syndrome (n=2). Mutation analysis for JAK2 V617F was performed on either bone marrow or peripheral blood cells using allele-specific polymerase chain reaction (AS-PCR) or sequencing and fluorescence resonance energy transfer (FRET) probes with MCA. Results: For the initial 30 samples, the detection rate of JAK2 V617F using MCA was comparable to the gold standard of the PCR sequencing methods. However, the turnaround times for MCA and PCR were 2 hours and 2 days, respectively. The detection rate of JAK2 V617F was 76.2{\%} for PV (homozygous in 14.3{\%}), 46.9{\%} for ET, 80{\%} for myelofibrosis (homozygous in 20{\%}), and 0{\%} for the other conditions. In PV, patients with homozygous JAK2 V617F presented with significantly longer disease durations than heterozygous patients. In ET, there were no differences in the clinical parameters of patients harboring JAK2 V617F compared with those with wild-type JAK2. Conclusion: Heterozygous and homozygous JAK2 V617F mutations can be identified using the rapid and reliable assay based on FRET probes and MCA. Detection of JAK2 V617F can be used to assist in the diagnosis of BCR/ABL-negative MPNs.",
keywords = "Essential thrombocythemia, JAK2, Melting curve analysis, Myeloproliferative neoplasms, Polycythemia vera",
author = "Ho, {Ching Liang} and Wu, {Yi Ying} and Hung, {Hsiu Man} and Chang, {Ping Ying} and Kao, {Wei You} and Chen, {Yeu Chin} and Chao, {Tsu Yi}",
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T1 - Rapid identification of heterozygous or homozygous JAK2 V617F mutations in myeloproliferative neoplasms using melting curve analysis

AU - Ho, Ching Liang

AU - Wu, Yi Ying

AU - Hung, Hsiu Man

AU - Chang, Ping Ying

AU - Kao, Wei You

AU - Chen, Yeu Chin

AU - Chao, Tsu Yi

PY - 2012/1

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N2 - Background/Purpose: The activating JAK2 mutation with a G-C to T-A transversion at codon 617 (JAK2 V617F) is associated with myeloproliferative neoplasms (MPNs), including polycythemia vera (PV), essential thrombocythemia (ET), and myelofibrosis. Here, we report a technical advance in the diagnosis of JAK2 V617F in MPNs by melting curve analysis (MCA). Methods: From January through December 2006, we prospectively enrolled 78 patients with PV (n=21), ET (n=32), myelofibrosis (n=5), secondary erythrocytosis (n=4), secondary thrombocytosis (n=2), acute myelocytic leukemia (n=4), chronic myelocytic leukemia (n=8), and myelodysplastic syndrome (n=2). Mutation analysis for JAK2 V617F was performed on either bone marrow or peripheral blood cells using allele-specific polymerase chain reaction (AS-PCR) or sequencing and fluorescence resonance energy transfer (FRET) probes with MCA. Results: For the initial 30 samples, the detection rate of JAK2 V617F using MCA was comparable to the gold standard of the PCR sequencing methods. However, the turnaround times for MCA and PCR were 2 hours and 2 days, respectively. The detection rate of JAK2 V617F was 76.2% for PV (homozygous in 14.3%), 46.9% for ET, 80% for myelofibrosis (homozygous in 20%), and 0% for the other conditions. In PV, patients with homozygous JAK2 V617F presented with significantly longer disease durations than heterozygous patients. In ET, there were no differences in the clinical parameters of patients harboring JAK2 V617F compared with those with wild-type JAK2. Conclusion: Heterozygous and homozygous JAK2 V617F mutations can be identified using the rapid and reliable assay based on FRET probes and MCA. Detection of JAK2 V617F can be used to assist in the diagnosis of BCR/ABL-negative MPNs.

AB - Background/Purpose: The activating JAK2 mutation with a G-C to T-A transversion at codon 617 (JAK2 V617F) is associated with myeloproliferative neoplasms (MPNs), including polycythemia vera (PV), essential thrombocythemia (ET), and myelofibrosis. Here, we report a technical advance in the diagnosis of JAK2 V617F in MPNs by melting curve analysis (MCA). Methods: From January through December 2006, we prospectively enrolled 78 patients with PV (n=21), ET (n=32), myelofibrosis (n=5), secondary erythrocytosis (n=4), secondary thrombocytosis (n=2), acute myelocytic leukemia (n=4), chronic myelocytic leukemia (n=8), and myelodysplastic syndrome (n=2). Mutation analysis for JAK2 V617F was performed on either bone marrow or peripheral blood cells using allele-specific polymerase chain reaction (AS-PCR) or sequencing and fluorescence resonance energy transfer (FRET) probes with MCA. Results: For the initial 30 samples, the detection rate of JAK2 V617F using MCA was comparable to the gold standard of the PCR sequencing methods. However, the turnaround times for MCA and PCR were 2 hours and 2 days, respectively. The detection rate of JAK2 V617F was 76.2% for PV (homozygous in 14.3%), 46.9% for ET, 80% for myelofibrosis (homozygous in 20%), and 0% for the other conditions. In PV, patients with homozygous JAK2 V617F presented with significantly longer disease durations than heterozygous patients. In ET, there were no differences in the clinical parameters of patients harboring JAK2 V617F compared with those with wild-type JAK2. Conclusion: Heterozygous and homozygous JAK2 V617F mutations can be identified using the rapid and reliable assay based on FRET probes and MCA. Detection of JAK2 V617F can be used to assist in the diagnosis of BCR/ABL-negative MPNs.

KW - Essential thrombocythemia

KW - JAK2

KW - Melting curve analysis

KW - Myeloproliferative neoplasms

KW - Polycythemia vera

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