Detection of low allele burden of JAK2 exon 12 mutations using TA-cloning in patients with erythrocytosis.
OBJECTIVE
Polycythemia vera (PV) is a clonal myeloproliferative neoplasia associated with the activation of the Janus-activating kinase 2 (JAK2) mutation. The aim of this study is to identify clonal expansion of exon 12 mutations.


METHODS
We performed DNA sequencing of the JAK2 exon 12 after TA-cloning in JAK2-V617F-negative and JAK2-V617F-positive PV patients.


CONCLUSIONS
We found clonal mutations (i.e. H538-K539delinsL and D544G) in 3 of 7 JAK2-V617F-negative PV patients, however, unlike JAK2-V617F, allele burden of JAK2 exon 12 mutation was low. Since allele-specific PCR is able to amplify only the limited region which contains known mutations with gain-of-function, we need to clarify the biological implications of unknown single nucleotide substitution of the JAK2 exon 12 with low clonal burden in erythrocytosis patients.
Ohyashiki
Junko H
JH
Intractable Disease Therapeutic Research Center, Tokyo Medical University, 6-7-1 Nishishinjuku, Shinjuku-ku, Tokyo 160-0023, Japan. junko@hh.iij4u.or.jp
Hisatomi
Hisashi
H
Shimizu
Syoko
S
Sugaya
Maki
M
Ohyashiki
Kazuma
K
D000328:Adult; D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D005091:Exons / Q000235:genetics*; D005260:Female; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D011086:Polycythemia / Q000235:genetics* / Q000473:pathology; D011087:Polycythemia Vera / Q000235:genetics* / Q000473:pathology; D011379:Prognosis; D015996:Survival Rate; D055815:Young Adult
C507924:JAK2 protein, human; D053614:Janus Kinase 2