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Kit for detecting 34 mutation sites of lung cancer based on MALDI-TOF-MS

A technology of mutation sites and kits, applied in recombinant DNA technology, microbial measurement/testing, DNA/RNA fragments, etc., can solve problems such as time-consuming, limited number of genes and sites, complex operations, etc.

Inactive Publication Date: 2018-08-17
GUANGZHOU DARUI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the limited number of genes and loci detected in a single test or the need for specialized bioinformatics analysis in the later stage, these methods have problems such as high cost, complicated operation, and long time consumption, which are not conducive to clinical large sample detection.

Method used

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  • Kit for detecting 34 mutation sites of lung cancer based on MALDI-TOF-MS
  • Kit for detecting 34 mutation sites of lung cancer based on MALDI-TOF-MS
  • Kit for detecting 34 mutation sites of lung cancer based on MALDI-TOF-MS

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Embodiment 1

[0134] 1. A kit for detecting 34 mutation sites in lung cancer based on MALDI-TOF-MS, said kit comprising: purified water, multiple PCR buffer, magnesium chloride buffer, dNTP mixture, multiple PCR enzyme, amplification primer mixture 1-2, phosphatase buffer, phospho-digesting enzyme, extension buffer, base extension reaction catalyzing enzyme, extension termination mixture A, T, C, G, extension primer mixture 1-8.

[0135] Wherein, the elongation termination mixture A, T, C, and G are all aqueous solutions of the mixture of four dideoxynucleotides, ddATP, ddTTP, ddCTP and ddGTP, wherein the ratio of the elongation termination mixture A is 1:4:4:4 , the ratio of extension-stop mixture T is 4:1:4:4, the ratio of extension-stop mixture C is 5:5:1:5, and the ratio of extension-stop mixture G is 3:3:3:1.

[0136] Multiplex PCR amplification reaction primers shown in SEQ ID NO: 1-14 constitute an amplification primer mixture 1;

[0137] The multiplex PCR amplification reaction pri...

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Abstract

The invention discloses a kit for detecting 34 mutation sites of lung cancer based on MALDI-TOF-MS. The kit comprises multiple PCR amplification reaction primers which are shown as SEQ ID NO:1-16, andsingle-base extension primers which are shown as SEQ ID NO:17-48. The kit provided by the invention can be used for simultaneously detecting 23 sites of an EGFR gene, 7 sites of a KRAS gene, 1 site of a BRAF gene and 3 sites of a PIK3CA gene. According to the kit provided by the invention, the 34 sites of the four genes can undergo target segment amplification in two holes and single-base extension in eight holes; and with the application of the kit provided by the invention, 34 site mutations of the four genes, which may exist in an abnormal sample, can be simultaneously detected and analyzed. The kit provided by the invention has important guiding significance for mastering related factors of a patient on drug susceptibility, conducting individualized treatment on the patient with the lung cancer and targeting to drug adaptability.

Description

technical field [0001] The invention relates to the technical field of lung cancer mutation site detection, and more specifically, to a kit for detecting 34 mutation sites in lung cancer based on MALDI-TOF-MS. Background technique [0002] Lung cancer is one of the most common malignant tumors in the world today. Its tumor morbidity and mortality rate rank first in men and second only to breast cancer in women. Chemotherapy is currently the main method for the treatment of advanced lung cancer, and it is also one of the preoperative and postoperative adjuvant treatments for patients. Clinically, some patients have poor chemotherapy effect and chemotherapy failure. The main reason is that the tumor cells are not sensitive or resistant to anticancer drugs. Therefore, it is particularly important to understand the relevant factors of patients' drug sensitivity for individualized treatment of lung cancer patients and guidance for the application of targeted drugs. [0003] It ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/156
Inventor 杨学习吴英松李明朱安娜李晓敏蔡晶晶
Owner GUANGZHOU DARUI BIOTECH
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