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JAK2 mutations

A G1920T, C1922T technology, applied in the field of cancer diagnosis and treatment, can solve problems such as inappropriate contact

Inactive Publication Date: 2010-12-15
QUEST DIAGNOSTICS INVESTMENTS INC
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Proper contacts between the two domains within the wild-type protein allow for proper kinase activation and regulation; however, the V617F mutation causes inappropriate contacts between the two domains, resulting in constitutive kinase activity within the mutant JAK2 protein

Method used

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Examples

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Embodiment 1

[0093] Example 1: Detection of JAK2 mutations.

[0094] Whole blood samples of 634 individuals with unknown MPD status were tested for JAK2 mutations in the region surrounding the codon encoding amino acid 617 of SEQ ID NO:3. 130 plasma samples from patients with confirmed MPD were also tested.

[0095] Total RNA was extracted from the mixture using the NucliSense Extraction Kit (bioMerieux Inc., Durham, NC) recommended by the manufacturer. A PCR primer pair was designed to amplify the region of the JAK2 gene including encoding amino acid 671. The primer sequences used for PCR and sequencing were as follows: JAK2-F (5'-GAC TAC GGT CAA CTG CAT GAA A-3') SEQ ID NO: 5 (corresponding to nucleotides 1776-1797 of SEQ ID NO: 1) and JAK2-R (5'-CCATGC CAA CTG TTT AGC AA-3') SEQ ID NO: 6 (corresponds to nucleotides 2029-2048 of SEQ ID NO: 1). One-step RT-PCR was performed using the SuperScript III One-Step RT-PCR System and Platinum Taq (Invitrogen, Carlsbad, CA) in a 25 μL reaction ...

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Abstract

Novel mutations in the JAK2 gene and JAK2 protein are disclosed. Methods and compositions useful for diagnosing or determining prognosis for an individual having a neoplastic diseases including, for example, myeloproliferative diseases are also disclosed.

Description

field of invention [0001] The present invention relates to the field of cancer diagnosis and therapy. Background of the invention [0002] The following discussion of the background of the invention is provided merely to aid the reader in understanding the invention and is not an admission that it describes or constitutes prior art to the invention. [0003] Some neoplastic diseases - including non-CML myeloproliferative disorders (MPDs) such as polycythemia vera (PV), essential thrombocythemia (ET) and chronic idiopathic myelofibrosis (IMF) and still as unclassified Myeloproliferative disease (MPD-NC) – characterized by an abnormal increase in blood cells. See, eg, Vainchenker and Constantinescu, Hematology (blood Hematology) (American Society of Hematology (American Society of Hematology)) , 195-200 (2005). This increase is usually triggered by spontaneous mutations in multipotent hematopoietic stem cells located in the bone marrow. ID Due to this mutation, stem cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N9/12
CPCC12N9/1205C12Q1/6886C12Q2600/156
Inventor M·阿尔比塔
Owner QUEST DIAGNOSTICS INVESTMENTS INC
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