Composition for detecting colorectal cancer hotspot gene mutation sites and using method of composition

A composition and hotspot technology, which can be applied in the fields of biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, etc., can solve problems such as low sensitivity, poor detection specificity, and inability to meet the needs of bowel cancer detection, etc. Achieve high sensitivity and specificity

Active Publication Date: 2014-10-15
JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, the sensitivity of these technologies is not high, the specificity of detection is poor, and the misdiagnosis rate is high, which cannot meet the needs of colorectal cancer detection.

Method used

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  • Composition for detecting colorectal cancer hotspot gene mutation sites and using method of composition
  • Composition for detecting colorectal cancer hotspot gene mutation sites and using method of composition
  • Composition for detecting colorectal cancer hotspot gene mutation sites and using method of composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1, screening of hotspot gene mutations in colorectal cancer patients

[0067] 1. Experimental materials

[0068] 1. Collect tissue samples and matching plasma samples from 20 patients with pathologically diagnosed bowel cancer. After the plasma samples were collected, they were immediately stored in a -80°C refrigerator.

[0069] 2. The purity of all primers should reach electrophoresis grade (PAGE) or HPLC grade, free of impurities. Provide the quality inspection certificate of the synthetic product issued by the synthesis institution, such as PAGE electrophoresis results or HPLC analysis patterns, which prove that there should be obvious single-peak PAGE or HPLC purification patterns after purification by PAGE or HPLC, and the concentration is 10ng / μl for use.

[0070] 3. All reagents were purchased from regular manufacturers: DNA polymerase (Roche), 10×PCR Buffer (Roche), MgCl2 (Roche), dNTP (TaKaRa), purified water.

[0071] 2. Main instruments: Roche Lig...

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Abstract

The invention discloses composition for detecting colorectal cancer hotspot gene mutation sites. The composition comprises primer sequences from SEQ NO:1 to SEQ NO:17 as well as block sequences from SEQ NO:18 to SEQ NO:21. The detecting method for mutations related to colorectal cancer has the advantages of high specificity, high sensitivity, low pollution, simplicity and rapidness in operation, safety and the like, detection results are better in accuracy and repeatability, the method is particularly suitable for detection of hotspot mutations of colorectal cancer driving genes from body fluids such as plasma and the like, real-time and non-invasive diagnosis, recurrence monitoring and curative effect evaluation can be performed on colorectal cancer patients, and the method has important values.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a composition for detecting hot spot mutations of intestinal cancer driving genes. Background technique [0002] Colon cancer is the general term for colorectal cancer and rectal cancer. The incidence of colorectal cancer from high to low is rectum, sigmoid colon, cecum, ascending colon, descending colon and transverse colon. In recent years, there is a trend of developing to the proximal end (right colon) . The incidence of bowel cancer is closely related to lifestyle, heredity, and colorectal adenoma. [0003] Studies have found that the occurrence of colorectal cancer is closely related to mutations in KRAS, BRAF, and PIK3CA hotspot genes. The K-ras gene is located on the short arm of chromosome 12, 35kb, and belongs to a member of the Ras gene family. Among the Ras genes, K-Ras has the greatest impact on human cancer. It acts like a molecular switch: normally it controls the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6848C12Q1/6858C12Q2531/113C12Q2527/107C12Q2549/126
Inventor 王弢李宗飞张利清张丽乐飚徐维琛
Owner JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD
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