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Gene detection kit for prognosing gastric cancer metastasis and use method of gene detection kit

A gene detection and kit technology, applied in the field of gene detection kits for predicting gastric cancer metastasis, can solve the problems of missed treatment opportunity, low sensitivity of metastases, high price, etc., to achieve prevention of tumor metastasis, long treatment time, and high repeatability Effect

Inactive Publication Date: 2017-05-10
苏州首度基因科技有限责任公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 1) Radionuclide bone scan (ECT): the diagnosis rate of gastric cancer bone metastases is relatively high, and metastases can be detected several months before bone metastases are found on X-ray plain film and CT, and the diagnosis of multiple gastric cancer bone metastases is false positive Rarely, but there is a certain false positive for the diagnosis of solitary gastric cancer bone metastasis;
[0010] 2) X-ray examination: it can be used as a basic examination method for the diagnosis of gastric cancer bone metastasis, it is helpful to detect the degree of destruction of bone marrow and cortical bone at the site of osteolytic bone metastasis, and it can also be used to confirm other imaging findings. Destroyed metastases have low sensitivity;
However, because of its high price, it cannot be widely used at present, and can only be used as a special inspection method.
[0019] At present, medical imaging is mainly used clinically for the clinical diagnosis of gastric cancer metastasis, but these methods also have some obvious shortcomings, mainly reflected in: the indicators are not objective enough, and cannot provide a clear judgment for micrometastases and critical conditions; The identified metastases are already in the middle and advanced stage for the patient, and the best treatment opportunity is missed, which is not conducive to the treatment of the patient

Method used

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  • Gene detection kit for prognosing gastric cancer metastasis and use method of gene detection kit
  • Gene detection kit for prognosing gastric cancer metastasis and use method of gene detection kit
  • Gene detection kit for prognosing gastric cancer metastasis and use method of gene detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] A gene detection kit for predicting gastric cancer metastasis, comprising:

[0050] - vacuum blood collection tube;

[0051] - Blood DNA extraction kit, which includes the following components: isopropanol, RNase, proteinase K and various enzyme reaction buffers, such as 10X ligation buffer, PCR buffer.

[0052] -Next-generation sequencing rapid DNA library construction kit, which includes 50 pre-designed gene probes, including high-risk gene probes and low-risk gene probes, of which:

[0053] - High-risk genes include: CDH1, CDH2, SNAIL, SLUG, MUC4, MUC6, PRSS3, USP6, MLH1, MSH2, MSH6, PMS2, TGFBR2, MMP2, MMP9, BRCA1, BRCA2, PALB2, ATM, ATR, MUTYH, EMSY, ERCC4 , RAD51, PARP1 and XRCC1;

[0054] - Low-risk genes include: ATRX, BRIP1, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, FANCO, FANCP, MDM2, MDM4, MLH1, NPM1, PP2R1A, PRKDC, RAD50, STAG2, XRCC5 and XRCC6. In addition, the next-generation sequencing rapid DNA library construction kit al...

Embodiment 2

[0056] The method for using the gene detection kit for predicting gastric cancer metastasis in Example 1 comprises the following steps:

[0057] (1) Blood collection: collect 5ml of blood from the patient, store it in a vacuum blood collection tube, and extract cfDNA within 72 hours;

[0058] (2) DNA extraction: Use a blood DNA extraction kit to extract free DNA in the blood, which contains a small amount of DNA released by gastric cancer tumor cells, including tumor cell-specific gene mutations;

[0059] (3) Library construction: use the next-generation sequencing rapid DNA library construction kit to build a library for cfDNA;

[0060] (4) Sequencing: The Illumina next-generation sequencing platform was used to sequence the DNA to obtain the full-length sequences of 50 genes;

[0061] (5) Gene mutation detection: After the sequencing results are off the machine, the gene mutation is analyzed through bioinformatics;

[0062] (6) Scoring mechanism: The mutations of high-risk...

Embodiment 3

[0072] Gastric cancer patients were diagnosed using the kits and methods in Examples 1-2.

[0073] Gastric cancer patient 1, named WLL, male, aged 61, clinically diagnosed as gastric cancer. After being tested by the kit of the present invention, the total score is 421, with a probability of over 50%, which is predicted to be a high risk of gastric cancer metastasis, and gastric cancer lymph node metastasis is detected after 10 months. The relevant mutations of the 50 genes detected and their specific scores are shown in Table 2-3:

[0074] Table 2 Number and score of high-risk gene mutations

[0075]

[0076] Table 3 Number and score of low-risk gene mutations

[0077]

[0078]

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Abstract

The invention relates to a gene detection kit for prognosing gastric cancer metastasis. The kit comprises a DNA library building kit, wherein the DNA library building kit comprises a high-risk gene probe and a low-risk gene probe; the high-risk gene probe comprises CDH1, CDH2, SNAIL, SLUG, MUC4, MUC6, PRSS3, USP6, MLH1, MSH2, MSH6, PMS2, TGFBR2, MMP2, MMP9, BRCA1, BRCA2, PALB2, ATM, ATR, MUTYH, EMSY, ERCC4, RAD51, PARP1 and XRCC1; the low-risk gene probe comprises ATRX, BRIP1, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, FANCO, FANCP, MDM2, MDM4, MLH1, NPM1, PP2R1A, PRKDC, RAD50, STAG2, XRCC5 and CRCC6. The invention further discloses a use method of the kit. The use method comprises the following steps: extracting cfDNA in a blood sample; building a library for the cfDNA through the DNA library building kit, and then sequencing the DNA to obtain a gene overall length sequence; carrying out gene mutation analysis on the gene overall length sequence.

Description

technical field [0001] The invention relates to the field of gene diagnosis kits, in particular to a gene detection kit for predicting gastric cancer metastasis and its application method. Background technique [0002] Cancer death is currently a major factor threatening human life and health. More than 90% of cancer patients die from tumor metastasis, and the survival period of cancer patients with metastasis generally does not exceed five years. In my country, the incidence of gastric cancer ranks second among malignant tumors, and the mortality rate ranks fourth. More than 90% of hospitalized cases of gastric cancer in my country are advanced gastric cancer. Even if radical treatment is performed, the five-year survival rate is still hovering around 30%. The main reason affecting the prognosis of gastric cancer is metastasis. Gastric cancer metastasis mostly occurs in the middle and late stage of patients, and only a small part occurs in the early stage of gastric cancer...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor 杨海平徐健唐元华
Owner 苏州首度基因科技有限责任公司
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