Methylated biomarker or combination thereof for detecting breast cancer and application
A biomarker and methylation technology, which can be used in biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc., and can solve problems such as increased breast cancer risk
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0054] 1. Plasma cfDNA or tissue extraction and methylation library construction
[0055] 1.1 Extraction of blood cfDNA or tissue DNA.
[0056] The specific operation steps of plasma DNA extraction were carried out according to the instructions of MagMAX™ Cell-Free DNA Isolation Kit of Life Company. The extraction steps of tissue DNA were carried out according to the DNeasy Blood&Tissue Kit operating instructions of QIAGEN;
[0057] 1.2 Conversion
[0058] The extracted cfDNA (10ng) or tissue DNA (50ng) was subjected to bisulfite conversion to deaminate the unmethylated cytosine in the DNA and convert it to uracil, while the methylated cytosine remained unchanged to obtain For the DNA after bisulfite conversion, the specific operation of the conversion was carried out according to the instructions of the EZ DNA Methylation-Lightning Kit of Zymo Research.
[0059] 1.3 End Repair
[0060] 1.3.1 Add the following reagents to the transformed 17ul sample for reaction:
[0061]...
Embodiment 2
[0125] This example discloses a combination of methylation-specific biomarkers for the diagnosis of breast cancer. The plasma samples of 74 cases of mammography and color Doppler ultrasound without abnormality and the plasma samples of 155 cases of breast cancer patients were used in the Different groups of methylation level differences screened out 66 methylation biomarkers related to breast cancer, and verified the differential methylation combination model in normal and breast cancer plasma samples with independent data sets, specifically Process: LASSO and Random Forest were used to perform marker screening and model building in the training set and test set, respectively, including 52 healthy individuals and 108 plasma cfDNA samples from breast cancer patients. 42 methylation markers were screened by the LASSO method, 50 methylation markers were screened by the random forest method, and finally a total of 26 methylation markers were further screened. Finally, in the indep...
Embodiment 3
[0131] Using the information of 26 methylation markers (SEQ ID NO.1-26) among them, 22 cases of mammography and color Doppler ultrasound without abnormal normal people and 47 cases of breast cancer patient plasma samples (among them, 0 phase 3 cases, I 12 cases in stage II, 22 cases in stage II, 8 cases in stage III, and 2 cases in stage IV) were subjected to random combination test, and logistic regression model was used for modeling and analysis, and four markers (cg23035715, cg26371731, cg04541368 and cg13973436) were further screened. The data analysis method is consistent with Example 2, Top1 (cg23035715) and Top1 / 2 / 3 / 4 (cg23035715, cg26371731, cg04541368 and cg13973436) at 98% specificity (22 / 22), the breast cancer of verification set 0 The detection sensitivities of stage I were 100% (3 / 3) and 100% (3 / 3), the detection sensitivities of breast cancer stage I were 91.67% (11 / 12) and 91.67% (11 / 12), and breast cancer II The detection sensitivities of stage III were 90.90% ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 


