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JAK2 gene mutation detection method and kit thereof

A JAK2E12F, JAK2E12R technology, applied in the field of JAK2 gene mutation detection method and its kit, can solve the problem of small detection range

Active Publication Date: 2014-07-23
苏州苏大赛尔免疫生物技术有限公司
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

These methods have the limitations of small detection range in practical application, and, from a clinical point of view, there is still a certain rate of missed detection.

Method used

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  • JAK2 gene mutation detection method and kit thereof
  • JAK2 gene mutation detection method and kit thereof
  • JAK2 gene mutation detection method and kit thereof

Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0029] The present invention will be further described below by taking a sample to be tested of a typical patient with polycythemia vera as an example.

[0030] (a) extract the DNA of the sample to be tested;

[0031] (b) using JAK2 gene exon No. 12 PCR amplification primers and JAK2 gene exon No. 14 PCR amplification primers to perform PCR amplification on the DNA of the sample to be tested in step (a) from the positive and negative directions;

[0032] (c) using the JAK2 gene exon 12 sequencing reaction primer and the JAK2 gene exon 14 sequencing reaction primer to sequence the DNA amplified in step (b) from both forward and reverse directions;

[0033] (d) Compare the sequencing results in step (c) with the standard JAK2 gene exon 12 and exon 14 from the positive and negative directions, and after sequence analysis: it is found that the exon 12 of the JAK2 gene (gene Accession number: NM_004972) ATGTTTCACAAAAATCAGAAATGAAGATTTGATA was inserted at the 2133th bp, a total of 3...

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Abstract

The invention relates to a JAK2 gene mutation detection method and a kit which have epoch-making meaning on classification and diagnosis of JAK2 mutation to myeloproliterative neoplasms, The invention aims at the problems of limitation of small detection scope on JAK2 mutation and existence of certain omission factor, so that the JAK2 gene mutation detection method and the kit with the characteristics of wide detection scope, low detection cost and high detection sensitivity can be provided, and can detect peripheral blood for the patients and marrow specimen for the patients who add heparin.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a JAK2 gene mutation detection method and a kit thereof. Background technique [0002] The JAK2 gene is located on the short arm of human chromosome 9 and has 25 exons. The molecular structure and function of JAK2 have been basically understood. James et al. (Nature.2005 Apr 28; 434(7037): 1144-1148.) experimented with JAK2 gene wild type and V617F mutant transfected cell lines in vitro and found that the wild type can activate STAT-mediated transcription process, while V617F Mutants do not have this feature. They further transplanted cells transfected with wild-type JAK2 gene and V617F mutant into mice, and found that mice transplanted with V617F mutant bone marrow could cause abnormal increase in blood cells. These in vivo and in vitro experiments showed that the V617F mutation of JAK2 gene can lead to the occurrence of MPN. The main site of JAK2 mutation is V617F in exon 14. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 费敏沈宏杰
Owner 苏州苏大赛尔免疫生物技术有限公司
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