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Method of detecting gene mutation

A gene and mutation technology, applied in the field of detection of gene insertion mutation, can solve the problem that the mutation cannot be detected accurately

Inactive Publication Date: 2008-08-13
KK TOSHIBA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This caused the problem that mutations could not be detected accurately

Method used

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  • Method of detecting gene mutation
  • Method of detecting gene mutation
  • Method of detecting gene mutation

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Embodiment

[0071]Specific examples detected by the method of the present invention are described in detail. CYP2D6*18, which has a 9-base gene insertion mutation in the human gene CYP2D6, was used as a target gene. The partial sequence of the CYP2D6 gene is shown in FIG. 5 . A 5.3 kbp CYP2D6 gene fragment was amplified by PCR from a heterozygous sample consisting of wild-type CYP2D6 and a mutant CYP2D6*18 insertion of 9 bases. This PCR product was introduced into pCR2.1 plasmid vector. Plasmids containing wild-type CYP2D6 or mutant CYP2D6*18 inserted with 9 bases were respectively obtained by cloning. LAMP amplification was performed using each plasmid as a template, respectively. This LAMP product is reacted with a probe immobilized on an electrode. Thereafter, the product bound to the probe is electrochemically detected.

[0072] (1) Amplification of target nucleic acid

[0073] [primer]

[0074] Primers used in the LAMP method were synthesized. Their sequence is shown below. ...

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Abstract

There is provided a method of detecting a gene insertion or deletion mutation, includes the steps of reacting a target nucleic acid with a wild-type probe and a mutant-type probe, detecting the amount of the target nucleic acid bound to the each probes, and comparing the amount of the target nucleic acid bound to the wild-type probe with the amount of the target nucleic acid bound to the mutant-type probe.

Description

Background of the invention [0001] The invention relates to a method for detecting gene insertion and deletion mutations. [0002] In recent years, techniques for detecting genetic polymorphisms have developed rapidly. A method using a probe immobilized on a substrate is one of these techniques. In this method, a target nucleic acid is detected by hybridization to a probe. [0003] In addition to single base substitutions, genetic polymorphisms also include insertion mutations, deletion mutations, and the like. There have been many reports on the detection of single base substitutions using the above method. However, there are few reports on detection of insertion mutations and deletion mutations using the above methods (see, for example, Nat Genet. 14, 441-7, 1996 and Nucleic Acid Res. 33e33, 2005). [0004] When a probe is used to detect an insertion mutation or a deletion mutation, the hybridization between the probe and the target nucleic acid may cause the mutation si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6827Y10T436/143333C12Q2563/113C12Q2545/101C12Q2527/107
Inventor 中村奈绪子
Owner KK TOSHIBA