Polymorphism detection probe, polymorphism detection method, method of evaluating drug efficacy, and reagent kit for polymorphism detection

A polymorphism detection, polymorphism technology, applied in biochemical equipment and methods, microbial determination/inspection, material excitation analysis, etc., can solve problems such as expensive, inaccessible, interstitial pneumonia, etc.

Inactive Publication Date: 2013-04-03
ARKRAY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the case where mutations exist in the above three places, it has been reported that the effect of anti-EGFR antibody drugs represented by cetuximab (which is used as a therapeutic drug for unresectable colorectal cancer) cannot be obtained (for example, Cancer Res. (2009 ); 69: (5). March1, 2009 (Non-Patent Document 3), BMC Cancer. (2011) Mar25; 11: 107 (Non-Patent Document 4))
[0004] Molecular targeted drugs such as cetuximab are very expensive and may have serious side effects such as interstitial pneumonia

Method used

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  • Polymorphism detection probe, polymorphism detection method, method of evaluating drug efficacy, and reagent kit for polymorphism detection
  • Polymorphism detection probe, polymorphism detection method, method of evaluating drug efficacy, and reagent kit for polymorphism detection
  • Polymorphism detection probe, polymorphism detection method, method of evaluating drug efficacy, and reagent kit for polymorphism detection

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0371] PCR and Tm analysis were performed using a fully automatic SNPs inspection device (trade name i-densy (trademark), manufactured by ARKRAY Corporation) and the inspection reagents listed in Table 5 below. The performance of the probes for polymorphism detection of each fluorescently-labeled oligonucleotide described in Table 6 was confirmed. In addition, the type of fluorescent dye is shown in parentheses at the 3' end of the probe. In addition, capital letters indicate bases showing polymorphisms. The following are the same.

[0372] As templates, the templates (oligo DNA) listed in Table 7 were used.

[0373] table 5

[0374] In 50 μL of reaction solution for Tm analysis:

1× Gene Taq universal buffer (manufactured by Nippon Gene)

lOμM probe

0.2μM

5 μM oligo DNA

0.2μM

[0375] Table 6

[0376] Forward probe

[0377]

[0378] Reverse probe

[0379]

[0380] Tm value is measured value

[0381] Table 7

[03...

Embodiment 2

[0391] The nucleic acid mixture shown in Table 8 (4 μl / reaction solution) or the purified human genome (100 copies / reaction solution) was used as a sample, and an automatic SNPs inspection device (trade name i-densy (trademark), manufactured by ARKRAY) was used. PCR and Tm analysis were performed. The recipe of the PCR reaction solution is shown in Table 9.

[0392] Table 8

[0393] sample:

[0394] Copy number of each plasmid in 4 μL of nucleic acid mixture:

[0395] plasmid name

Wt sample

Mt Sample (1)

Mt samples (2)

exon9Wt DNA

1000

14000

18940

G1624A mt DNA

0

1000

60

G1633A mt DNA

0

5000

1000

exon20Wt DNA

1000

9700

9970

A3140G mt DNA

0

300

30

[0396] Table 9

[0397] PCR reaction solution

[0398] In 50 μL of PCR reaction solution:

[0399]

[0400] In PCR, after treating at 95° C. for 60 seconds, one cycle of 1 ...

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Abstract

The invention relates to a polymorphism detection probe, a polymorphism detection method, a method of evaluating drug efficacy, and a reagent kit for?polymorphism detection, concretely the invention provides the polymorphism detection probe for highly-sensitively and simply detecting a polymorphism in the PIK3 CA gene. The probe for detecting the polymorphism in the PIK3 CA gene according to the present invention is at least one fluorescently labeled oligonucleotide selected from P1 to P11' oligonucleotides.

Description

technical field [0001] The present invention relates to a probe for detecting polymorphism, a method for detecting polymorphism, a method for judging drug efficacy and a kit for detecting polymorphism. Background technique [0002] The PIK3CA gene encodes a PI3 kinase that regulates signaling pathways that play important functions in tumorigenesis. It is known that mutations of the PIK3CA gene are common in tumor tissues, especially G1624A (E542K) mutation, G1633A (E545K) mutation and A3140G (H1047R) mutation are found at high frequency (for example, see Science. (2004) Apr23; 304 (5670) :554 (Non-Patent Document 1) and Oncogene. (2005) Feb 17; 24(8):1477-80 (Non-Patent Document 2)). [0003] In the case where mutations exist in the above three places, it has been reported that the effect of anti-EGFR antibody drugs represented by cetuximab (which is used as a therapeutic drug for unresectable colorectal cancer) cannot be obtained (for example, Cancer Res. (2009 ); 69: (5)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11G01N21/64
CPCC12Q2600/156C12Q1/6886C12Q1/6827C12Q2527/107C12Q2563/107C12Q2565/107C12Q1/6876C12Q2600/106
Inventor 小森真理子
Owner ARKRAY INC
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