Colorectal cancer marker and application thereof

A technology of colorectal cancer and markers, which is applied in the fields of colorectal cancer markers, kits, colorectal cancer detection, primer pairs, and nucleic acid probes. And other issues

Active Publication Date: 2020-03-03
SHENZHEN HUADA GENE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional screening methods for colorectal cancer are mainly colonoscopy, in addition to biopsy and exfoliative cytology. , few clinical applications
Therefore, if there are no relevant symptoms, many people are not willing to do early screening for colorectal cancer

Method used

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  • Colorectal cancer marker and application thereof
  • Colorectal cancer marker and application thereof
  • Colorectal cancer marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The MGWAS method was used to search for markers of colorectal cancer intestinal flora (this method is well known in the art and described in detail in the literature), and the specific sequence of Fusobacterium nucleatum was found, as shown in Table 1 below. Among them, the MGWAS (genome-wide association analysis) method uses natural populations and artificial populations as research materials, and conducts genome-wide association analysis of metabolomes based on resequencing information and combined with metabolome results. References for the experimental process: Jun Yu, QiangFeng eal.,. Metagenomic analysis of faecal microbiome as a tool towards targeted non-invasive biomarkers for colorectal cancer. Gut 2015; 0:1–9. doi:10.1136 / gutjnl-2015-309800, according to It needs to be incorporated in whole or in part into this application.

[0043] Table 1 Specific gene sequence

[0044]

[0045]

[0046]

[0047]

[0048] Wherein, the base N in the sequence SEQ ...

Embodiment 2

[0050] The oligo7 software was used to design primers for the specific sequences of the specific bacterial groups screened. On the basis of meeting the important principles as much as possible, the amplification products of the upper and lower primers were required to be 70bp-200bp, and it was estimated that 3-5 sets of primers were designed for each gene.

[0051] For the specific sequences obtained by screening in Example 1, the primer sequences shown below were designed for the specific amplification of each specific sequence, and the designed primer sequences are shown in the following table:

[0052] Table 2 Primer Sequence

[0053]

[0054]

[0055] For the primers that are obtained by design, carry out primer test: promptly utilize common PCR to amplify, agarose gel electrophoresis detects the amplified band, finds through testing: the purpose band size is consistent with expectation and band is single, illustrates and utilizes the present invention to implement T...

Embodiment 3

[0057] The embodiment of the present invention is mainly based on the QPCR technology to test the fecal DNA of colorectal cancer patients and normal people, and the results shown are only the results provided by the best primers provided in Example 2. The stool DNA of 16 colorectal cancer patients and 16 normal people were used for QPCR detection (the reagent used was: SYBR Premix Ex Taq(TM) II (Perfect Real Time), product number: DRR081A). Among them, SYBR Premix Ex Taq(TM) II is a special reagent for Real Time PCR (real-time fluorescent quantitative PCR) using the SYBRGreen I mosaic method. Double-stranded DNA is generated by PCR reaction, and SYBR Green I can combine with double-stranded DNA to emit fluorescence. By detecting the intensity of the fluorescent signal in the PCR reaction solution, the target gene can be accurately quantified. Specifically, in the PCR reaction system, an excess of SYBR fluorescent dye is added, and after the SYBR fluorescent dye is specifically...

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Abstract

The invention relates to the field of gene detection, specifically to a colorectal cancer marker and application thereof. The marker comprises at least one of the following sequences: SEQ ID NO: 1 toSEQ ID NO: 5; a nucleic acid sequence derived from Fusobacterium nucleatum and having more than 95% of homology with any one of the SEQ ID NO: 1 to SEQ ID NO: 5. The invention also provides a primer pair, a nucleic acid probe, a construct, a kit and a colorectal cancer detection system for subjects to be tested. The marker provided by the invention can realize the rapid detection of colorectal cancer.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a colorectal cancer biomarker and its application, in particular to a colorectal cancer marker, a primer pair, a nucleic acid probe, a kit and a colorectal cancer detection method. Background technique [0002] According to statistics, colorectal cancer (CRC) ranks third in incidence and fourth in mortality among various cancers, and nearly 55% of the cases occur in developed countries. The incidence rate is low in people younger than 40 years old, but the incidence rate gradually increases with age. Colorectal cancer is also one of the most preventable tumors at present. The medical profession believes that if detected early, bowel cancer is the most curable cancer. The traditional screening methods for colorectal cancer are mainly colonoscopy, in addition to biopsy and exfoliative cytology. , less clinical application. Therefore, many people are not willing to do early screenin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851C12Q1/6886C12N15/85C12N15/11C12M1/34
CPCC12Q1/6851C12Q1/6886C12N15/85C12Q2600/118C12Q2563/107C12Q2531/113
Inventor 李南南揭著业易吉吴逵朱师达贾慧珏仝欣罗慧娟陈小芳
Owner SHENZHEN HUADA GENE INST
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