Colorectal cancer markers and their applications
A colorectal cancer and marker technology, applied in primer pairs, chips, kits and their applications, colorectal cancer markers, nucleic acid probe fields, can solve the problems of large trauma and little diagnostic value of carcinoembryonic antigen
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0038] Example 1 Screening and verification of biomarkers
[0039] A total of 193 stool samples were collected from Southwest Hospital in Chongqing, China, including 98 colorectal cancer patient samples (68 male samples, 30 female samples) and 95 healthy control samples (49 male samples, 46 female samples) . We performed metagenomic testing on 52 colon cancer patient samples and 55 healthy control samples (from healthy family members with similar dietary structure, living habits, and living environment) to detect colorectal cancer. Differences in gut microbiota between patients and healthy individuals. In the process of testing the above samples, we identified 30 microbial gene fragments associated with colorectal cancer, of which 26 gene fragments were increased in samples from patients with colorectal cancer, and 4 gene fragments were found in healthy control samples increased in ( figure 1 ). figure 2 Receiver operating characteristic (ROC) curve showed that 30 candida...
Embodiment 2
[0056] Example 2 Primer Screening
[0057] For the specific sequences of biomarkers obtained by screening in Example 1, design specific primers, and carry out primer testing for the specific primers designed for each specific sequence: that is, by ordinary PCR amplification, agarose gel electrophoresis is used to detect its stripes. If the size of the target band is the same as expected and the band is single, it indicates that the specificity of the primer is good. After testing, it was found that the primers shown in Table 2 could be used to amplify the specific sequences screened in Example 1, the size of the target band was the same as expected, the band was single, and the amplification specificity was good.
[0058] Table 2 Specific primer sequences
[0059]
Embodiment 3
[0060] Example 3 Colorectal cancer detection application
[0061] This example is mainly based on real-time fluorescence quantitative PCR technology to test the fecal microflora DNA of colorectal cancer patients and normal people, and the results shown are the results provided by the best primers provided in Example 2.
[0062] 1. Sample collection
[0063] Sampling swabs were used to collect the central part of feces from 30 colorectal cancer patients and 30 normal people, respectively, and store them in a fecal collection tube containing buffer solution. The feces are mixed evenly.
[0064] Storage and transportation of specimens: Store at room temperature or refrigerated at 4°C. Long-distance transportation of specimens before refrigeration can be transported at room temperature, and dry ice is required for long-distance transportation of specimens after refrigeration.
[0065] 2. Nucleic acid extraction
[0066] Use the fecal gene extraction kit QIAamp DNA Stool Mini K...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More