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Method for measuring expression level of abl1 t315i mutation

A technique for expression level and measurement object, which is applied in the field of measurement of the expression level of ABL1 T315I mutation, and can solve problems such as insufficient purpose and lack of quantification.

Pending Publication Date: 2020-04-10
OTSUKA PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, currently developed assays for the T315I mutation lack sufficient quantification to be adequate for these purposes.

Method used

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  • Method for measuring expression level of abl1 t315i mutation
  • Method for measuring expression level of abl1 t315i mutation
  • Method for measuring expression level of abl1 t315i mutation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0162] Primer and probe design

[0163] The following primer sets and probes were designed and synthesized.

[0164] Among the fluorescently labeled probes, the 5' end of the probe was labeled with HEX (6-carboxyfluorescein), and the 3' end of the probe was labeled with Iowa Black FQ (Integrated DNA technologies) as a quencher dye.

[0165] Details of the primers and probes used in this example are shown in Table 2.

[0166] [Table 2]

[0167]

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PUM

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Abstract

This disclosure provides a method that contains (1) a step for reverse transcribing a target RNA specimen in the presence of a modified nucleic acid having a base sequence complementary to a region that contains the T315I mutation position of wild-type ABL1 mRNA, using, in the same container, (a) a reverse primer that binds to a region downstream from the T315I mutation position of ABL1 mRNA and (b) a reverse primer that binds to a region upstream from the T315I mutation position of ABL1 mRNA, and (2) a step for determining the expression level of the ABL1 T315I mutation on the basis of the ratio of the reverse transcription product due to the primer (a) with respect to the reverse transcription product due to the primer (b). This disclosure also provides a kit for this method.

Description

technical field [0001] This patent application claims priority based on Japanese Patent Application No. 2017-087578, which is incorporated herein by reference in its entirety. [0002] The present application relates to a method for determining the expression level of the ABL1 T315I mutation, or a kit for the method. Background technique [0003] The BCR-ABL1 fusion gene is a chromosomal abnormality seen in Chronic Myelogenous Leukemia (CML) and Philadelphia chromosome positive Acute Lymphocytic Leukemia (Ph-positive ALL). Due to the translocation t(9;22) between chromosome 9 and chromosome 22, the ABL1 gene located on the q34 band of chromosome 9 is fused with the BCR gene located on the q11 band of chromosome 22 to form the fusion gene. The chimeric protein BCR-ABL1 encoded by the BCR-ABL1 fusion gene has tyrosine kinase activity, which can stably stimulate cell proliferation signals and inhibit apoptosis inhibition to make hematopoietic stem cells proliferate indefinitel...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/62
CPCC12N15/62C12Q1/6883C12Q2600/158
Inventor 葛城肃典田中秀明伊藤隆太古贺大辅
Owner OTSUKA PHARM CO LTD
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