Kit and method for detecting human KRAS gene mutation

A kit and human detection technology, applied in the field of molecular biology, can solve the problems of poor clinical prognosis and large differences in the efficacy of EGFR-TKI, and achieve the effects of less contamination, simple and reasonable structural design, and convenient use

Inactive Publication Date: 2014-06-25
瑞希基因科技(北京)股份有限公司
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Problems solved by technology

[0011] Three EGFR-TKIs (gefitinib / Iressa, erlotinib / Tarceva, and Camistat) are currently the main drugs for molecularly targeted therapy in NSCLC, but in different patients EGFR- Efficacy of TKIs varies widely
A large number of clinical trials have found that, in addition to EGFR mutations, KRAS mutations can help NSCLC patients better predict the efficacy of gefitinib; NSCLC patients with KRAS mutations have poorer clinical prognosis after combined treatment with erlotinib

Method used

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  • Kit and method for detecting human KRAS gene mutation
  • Kit and method for detecting human KRAS gene mutation
  • Kit and method for detecting human KRAS gene mutation

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Embodiment 1

[0050] 1) Amplification primers:

[0051] KARS codon 12, 13F: 5'-AGGCCTGCTGAAAATGACTGAAT-3' (SEQ ID NO.1);

[0052] KARS codon 12, 13R: 5'-GTTGGATCATATTCGTCCACAAA-3' (SEQ ID NO.2), 5' terminal biotin labeling;

[0053] KARS codon 61F: 5'-CAGACTGTGTTTCTCCCCTTCTCAGGATT-3' (SEQ ID NO. 3);

[0054] KARS codon 61R: 5′-AAGAAAGCCCTCCCCAGTCC-3′ (SEQ ID NO.4), 5′ terminal biotin-labeled;

[0055] 2) Sequencing primers:

[0056] KARS codon 12, 13: 5'-AAACTTGTGGTAGTTGGAGC-3' (SEQ ID NO.5);

[0057] KARS codon 61: 5'-TTGGATATTCTCGACACAGCAGGT-3' (SEQ ID NO. 6).

[0058] 1. Extract sample DNA:

[0059] 1. Sample Processing

[0060] a. Paraffin section: Take 5-8 paraffin sections (5-10μm thick, 1×1cm2 size).

[0061] b. Paraffin block: scrape about 30 mg of tissue sample with a scalpel (remove excess paraffin as much as possible).

[0062] NOTE: If the sample surface is exposed to air, discard the first 2-3 scraps.

[0063] c. Samples in fixative solutions such as formalin: take 30m...

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Abstract

The invention relates to a kit and a method for detecting human KRAS gene mutation. The kit comprises two pairs of amplification primers and two sequencing primers. The kit for detecting human KRAS gene mutation, which is disclosed by the invention, is designed completely based on a novel experiment method, is simple and reasonable in structural design, is convenient to use, and has the advantages of high sensitivity, high specificity, short time consumption and the like when being used for detecting KRAS gene mutation.

Description

technical field [0001] The invention relates to a kit for detecting human KRAS gene mutation and a detection method thereof, belonging to the field of molecular biology. Background technique [0002] 1) Clinical significance of KRAS gene mutation in colorectal cancer [0003] In recent years, with the continuous improvement of people's living standards, changes in eating habits and dietary structure, and population aging, the incidence and mortality of colorectal cancer (CRC) in my country have maintained an upward trend. Among them, the incidence of colon cancer increased significantly. Most patients are already in the middle and late stages when they are discovered. The incidence of colorectal cancer is 221,000, the mortality rate is 110,000, and the 5-year survival rate is 63.7%. [0004] KRAS is a key downstream regulator in the EGFR (epidermal growth factor receptor) signaling pathway, involved in the regulation of cell growth, and plays an important role in the proc...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor王彤陈大方
Owner瑞希基因科技(北京)股份有限公司