Method and kit for screening gastric cancer related gene mutation

A technology of kits and detection kits, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve problems such as deviations and use limitations, and achieve the effects of low requirements, low cost, and short detection time

Pending Publication Date: 2021-11-02
谢小冬
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, there will be a large deviation in the prediction of the occurrence and development of gastric cancer, and there will be great limitations in its use.

Method used

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  • Method and kit for screening gastric cancer related gene mutation
  • Method and kit for screening gastric cancer related gene mutation
  • Method and kit for screening gastric cancer related gene mutation

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Embodiment 1

[0048] Such as Figure 1-4 As shown, a method and kit for screening gastric cancer-related gene mutations according to the present invention include;

[0049] 1) Capture: Perform the first round of specific multiplex PCR reaction, and use 61 pairs of primers to amplify the sample DNA. The 61 pairs of PCR primers are reacted in T1 test tubes, and the 61 pairs of test tube primers are SEQ ID NO.1-SEQ ID NO .61;

[0050] 2) Library construction: carry out the second round of linker PCR reaction, use the linker sequences as IGT-I5 Index and IGT-I7Index, amplify the amplified product obtained in step 1), and obtain a multiplex PCR library;

[0051] 3) Quantitative and quality detection: measure the library concentration of the multiple PCR library obtained in step 2), and detect the length and purity of the library fragments;

[0052] 4) Sequencing: Sequencing the multiplex PCR library obtained in step 2);

[0053] 5) Analysis: analyze the sequencing results obtained in step 4),...

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Abstract

The invention discloses a method and kit for screening gastric cancer related gene mutation, and belongs to the technical field of gene mutation detection. The method and the kit use two sets of PCR primers: the first set of PCR primers comprises 61 pairs of PCR primers, the first set of PCR primers comprises 61 pairs of PCR primers (SEQ ID NO.1-SEQ ID NO.61), and the 61 pairs of PCR primers cover 63 sites of gastric cancer related gene mutation; and the second set of primers are universal primers I GT-I5I ndex (SEQI D NO.17) and I GT-I7I ndex (SEQ ID NO.18). The kit comprises a PCR (Polymerase Chain Reaction) primer combination, an I ndex linker sequence, an I GT-EM808 polymerase mixture, an amplification buffer enhancer NB, a YF magnetic bead rinsing buffer solution B, a reagent M and the like. Compared with the prior art, the detection method for detecting the gastric cancer related gene mutation disclosed by the invention has the advantages that the required quantity of a sample to be detected is low; an extremely low DNA sample in plasma and serum can be analyzed; through the design of the carpet type primer, mutation detection of a full coding region can be realized, known mutation can be analyzed, and new mutation can be found; the cost is low; the sensitivity is high; and the detection time is short.

Description

technical field [0001] The invention belongs to the technical field of gene mutation detection, and in particular relates to a method and a kit for screening gastric cancer-related gene mutations. Background technique [0002] Gastric cancer is one of the most common malignant tumors in the world. The annual incidence of gastric cancer worldwide is 17.6 / 100,000, ranking second in the mortality rate of malignant tumors. Studies have shown that the incidence of gastric cancer can vary by 7-10 times between high-incidence areas and low-incidence areas, and there are significant differences between regions. Gastric cancer mortality rates in Japan rank first in the world, followed by Chile and Costa Rica, and this trend is true for both men and women. Gastric cancer mortality rates are lower in New Zealand, Denmark, and Australia, the lowest in the United States, and about seven times higher in Japan. The incidence of gastric cancer in Japan has decreased in recent years, but i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6869
CPCC12Q1/6886C12Q1/6869C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2525/191C12Q2535/122
Inventor 谢小冬赵龑飞马青龙
Owner 谢小冬
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