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Kit for detecting PIK3CA gene mutation of colorectal cancer patient

A colorectal cancer and kit technology, applied in the field of biomedicine, can solve the problems of inconvenient operation and low detection sensitivity, achieve high sensitivity, reduce false negative results, and avoid false positive results

Inactive Publication Date: 2017-11-21
安徽安龙基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a test kit for detecting PIK3CA gene mutation in colorectal cancer patients in order to overcome the defects of low detection sensitivity and inconvenient operation in the prior art

Method used

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  • Kit for detecting PIK3CA gene mutation of colorectal cancer patient
  • Kit for detecting PIK3CA gene mutation of colorectal cancer patient
  • Kit for detecting PIK3CA gene mutation of colorectal cancer patient

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The method for detecting 5 gene mutations of PIK3CA by real-time fluorescent PCR of the present invention comprises the following steps:

[0042] (1) Nucleic acid extraction

[0043] Taking paraffin-embedded tissue as an example, extract genomic DNA from paraffin-embedded tissue according to the recommended QIAamp DNA FFPE Tissue Kit instructions.

[0044] (2) Preparation of PCR reaction system (25ul)

[0045] Reagent name

Amount added

PCR reaction solution

16ul

Enzyme 1

2ul

Primer Probe Mix

2ul

template DNA

5ul

[0046] The template DNA includes sample DNA and negative and positive control DNA.

[0047] (3) PCR amplification

[0048] Real-time PCR amplification conditions are: 50°C for 2min, 1 cycle; 95°C for 5min; 95°C for 10s, 55°C for 15s, 72°C for 30s, 5 cycles; 95°C for 10s, 60°C for 45s (collect FAM and HEX fluorescence signals) , 40 cycles.

[0049] (4) Result judgment

[0050] 1) In addition to the...

Embodiment 2

[0059] Using the present invention to detect 10 cases of paraffin-embedded tissue samples from PIK3CA gene mutation-positive patients, and compared with the sequencing results, the results: 5 cases were detected positive for H1047R, 3 cases were positive for E542K, and 2 cases were positive for E545K, and the coincidence rate with the sequencing method was 100%. .

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Abstract

The invention discloses a kit for detecting PIK3CA gene mutation of a colorectal cancer patient. The kit is composed of the following components of 192 ul of a PCR reaction solution, 1-24 ul of an enzyme system, 24 ul of an EC / IC primer probe, 24 ul of an E542K / IC primer probe, 24 ul of an E545K / IC primer probe, 24 ul of an E542D / IC primer probe, 24 ul of an H1047R / IC primer probe, 24 ul of an H1047L / IC primer probe, 200 ul of a positive control agent and 500 ul of a negative control agent. The kit for detecting PIK3CA gene mutation of the colorectal cancer patient has the advantages that the detection sensitivity is high, and detection is convenient.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a kit for detecting PIK3CA gene mutation in patients with colorectal cancer. Background technique [0002] The PIK3CA gene was detected by Volinia using in situ hybridization in 1994. It is located at 3q26.3, 34kb in length, contains 21 exons, and encodes 1 068 amino acids. This group of amino acids produces a group of 124kD proteins . PIK3CA encodes the p110 catalytic subunit of class I phosphatidylinositol 3-kinases (PI3Ks), namely PI3Kp110a, and about 4 / 5 of the mutations in PIK3CA occur in the helical region (exon9) and the kinase region (exon20 ) of these two hot spots. [0003] Colorectal cancer is one of the most common malignant tumors in the world. In China, the morbidity and mortality of colorectal cancer rank among the top ten of all tumors. In 2014, the incidence and mortality were 16 / 100,000 and 7.5 / 100,000, respectively. The morbidity and mortality of colorectal cance...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q1/6848C12Q2600/156C12Q2545/101C12Q2545/113C12Q2531/113
Inventor 韦玉军李航刘文干苏军吴远航
Owner 安徽安龙基因科技有限公司
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