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Nucleotide sequence, method and kit for detecting exons 12, 13 mutation of human K-ras gene

A nucleotide sequence and exon technology, applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc. Aiming at advanced background problems

Active Publication Date: 2011-04-13
上海源奇生物医药科技有限公司 +1
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AI Technical Summary

Problems solved by technology

Taqman probes are the earliest fluorescent probes with the highest penetration rate. At present, almost all fluorescent PCR kits used in the domestic market use this type of probe, but due to the high background of this type of probe and the inability to identify a single nucleus The difference in nucleotides is not conducive to the mutation detection of Kras gene

Method used

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  • Nucleotide sequence, method and kit for detecting exons 12, 13 mutation of human K-ras gene
  • Nucleotide sequence, method and kit for detecting exons 12, 13 mutation of human K-ras gene
  • Nucleotide sequence, method and kit for detecting exons 12, 13 mutation of human K-ras gene

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Embodiment 1

[0070] 1. The raw material that the present invention adopts

[0071] 1.1 Primers and probes

[0072] Primers and probes were synthesized by Shanghai Chaoshi Biotechnology Co., Ltd.

[0073] 1.2 Other materials

[0074] Purified water complies with the requirements of "Quality Control Standards for Main Raw and Auxiliary Materials of Biological Products in China"; Taq enzyme, dNTP (dATP:dCTP:dGTP:dUTP=25mM:25mM:25mM:25mM), and UNG enzyme are provided by Promega. All chemical reagents are analytically pure or above, and all reagents have passed the inspection.

[0075] 1.3 Reference substance

[0076] 1.3.1 Negative control substance: purified water; the substitution test is qualified.

[0077] 1.3.2 Positive control substance: known Kras wild-type and mutant plasmid DNA solutions.

[0078] 2. Detection method

[0079] 2.1 Specimen extraction: Genomic DNA in human colon cancer tissue was extracted with a genomic DNA extraction kit.

[0080] 2.2 Preparation of PCR reactio...

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Abstract

The invention relates to a nucleotide sequence, a method and a kit for detecting exons 12, 13 mutation of a human K-ras gene. The nucleotide sequence for detecting the exons 12, 13 mutation of the human K-ras gene comprises any one nucleotide sequence shown from SEQ ID NO:1 to SEQ ID NO:10, wherein two ends of the nucleotide sequence shown in the SEQ ID NO:3 are labeled with JUP (Junction Plakoglobin) fluorescence groups, and two ends of the nucleotide sequences shown in the SEQ ID NO:4 to the SEQ ID NO:10 are labeled with MAR fluorescence groups. The kit of the invention can be used for instructing the clinical medication of Erbitux and other EGFR (Epithelial Growth Factor Receptor) antibody medicines.

Description

technical field [0001] The invention relates to a nucleotide sequence for detecting mutations of exons 12 and 13 of human K-ras gene, a method for detecting mutations of exons 12 and 13 of human K-ras gene and a kit. Background technique [0002] Targeted therapy has become a new hot spot in the field of cancer in recent years. Compared with the effect of traditional chemotherapy, targeted drugs can significantly prolong the progression-free survival (PFS) and overall survival (OS) of patients, while reducing the adverse reactions of drugs. The US FDA has successfully approved the following drugs for the treatment of the above tumors, such as: Trastuzumab (Herceptin) for breast cancer, Gefitinib (Iressa) and Erlotinib (Tarceva) for non-small cell lung cancer, Cetuximab (Love) Bidu) for the treatment of colorectal cancer, Sunitinib and Sorafenib for the treatment of renal cell carcinoma. [0003] Since targeted diagnosis is required before targeted therapy, how to accurately...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 熊慧
Owner 上海源奇生物医药科技有限公司
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