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
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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
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[0078]
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