Method and system for evaluating colorectal cancer metastasis and recurrence risk and dynamically monitoring based on methyl marker combination
A colorectal cancer and risk monitoring technology, applied in the field of bioinformatics, can solve problems such as poor sensitivity and stability, and achieve the effects of improving survival rate and quality of life, improving usability, and excellent sensitivity
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Embodiment 1
[0054] The collection of embodiment 1 tissue and plasma sample:
[0055] (1) 191 formalin-fixed and paraffin embedded tissues (Formaldehyde-fixed and paraffin embedded tissues, FFEP) samples and 263 plasma samples were collected from the Department of Pathology and General Surgery of Southern Hospital and Southern Theater General Hospital.
[0056] FFEP tissue: 52 cases of normal intestinal mucosa (outer intestinal mucosa >1cm around the tumor) and 138 cases of colorectal cancer tissue (27 cases of stage I, 30 cases of stage II, 33 cases of stage III, 35 cases of primary tumor of stage IV, 35 cases of primary tumor of stage IV 40 cases of distant metastases); Among them, 25 cases of paired samples of stage IV primary tumors and metastases. There were 86 male patients and 53 female patients, with an average age of 58 (25-79 years old). Select a tissue block with tumor tissue accounting for more than 30%, slice the thickness of 5-6 μm, cut 6-8 slices, remove the slice and stick...
Embodiment 2
[0061] Embodiment 2 bisulfite conversion
[0062] Sulphite Conversion Sulphite conversion was performed according to the instructions of the EZ DNA Methylation Modification Kit ((Cat#D5031, Zymo Research).
Embodiment 3
[0063] Example 3 Baseline Medical IRIS TM Targeted Methylation Sequencing
[0064] Anchor IRIS TM Pre-library construction using Anchor Dx EpiVisio TM Methylation Pre-Library Construction Kit (Anchor Dx, Cat#A0UX00019) and EpiVisio TM The indexing PCR kit (Anchor Dx, Cat#A2DX00025) was used for pre-library construction. Subsequent purification of the amplified pre-library using IPB1 magnetic beads, Qubit TM The dsDNA HS assay kit detects the pre-library concentration. If the DNA content in the pre-library exceeds 400ng, further targeted enrichment can be performed.
[0065] Anchor IRIS TM Target Enrichment Benchmark Medical Company EpiVision TM The Targeted DNA Enrichment Kit (AnchorDx, Cat#A0UX00031) performs efficient liquid-phase capture of the DNA in the pre-library to enrich the targeted region. A 10K methylation panel containing 9921 cancer-specific methylated preselected regions was selected to obtain the enriched final library. The 10K panel is based on the ...
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