Primers, kits, models and construction methods for methylation of cervical cancer-related genes

A construction method and methylation technology, applied in biochemical equipment and methods, biological systems, combinatorial chemistry, etc., can solve problems such as inaccurate assessment of cervical cancer methylation levels, achieve high sensitivity and specificity, and reduce incidence. Rate and mortality, the effect of high sequencing depth

Active Publication Date: 2022-04-19
北京迈基诺基因科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Aiming at the deficiencies of the above-mentioned prior art, the problem to be solved by the present invention is: how to provide a primer, kit, model and construction method for the methylation of cervical cancer-related genes, so as to solve the inaccurate assessment of the methylation level of cervical cancer The problem

Method used

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  • Primers, kits, models and construction methods for methylation of cervical cancer-related genes
  • Primers, kits, models and construction methods for methylation of cervical cancer-related genes
  • Primers, kits, models and construction methods for methylation of cervical cancer-related genes

Examples

Experimental program
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Effect test

Embodiment 1

[0045] 1. DNA extraction from cervical cells

[0046] DNA extraction from cervical scraping cells: Take 300 μL of preservation solution to preserve the cells and extract the genomic DNA according to the instructions of the Universal Column Genome Extraction Kit (Kangwei, CWY004) and then measure the concentration with Qubit.

[0047] Genomic DNA extraction from paraffin tissue or paraffin section: Take 10 μm thick paraffin block or 8-10 paraffin section samples, extract genomic DNA according to the instructions of GeneRead DNA FFPE Kit (Qiagen, 180134) and then measure the concentration with Qubit.

[0048] Genomic DNA extraction from fresh tissue: Take 25 mg of fresh tissue to extract genomic DNA according to the instructions of the Universal Column Genome Extraction Kit (Kangwei, CWY004) and then measure the concentration with Qubit.

[0049] 2. Methylation treatment

[0050] DNA obtained from cervical cells was methylated using the Epitect Plus DNA Plus DNA Bisufite Kit (Q...

Embodiment 2

[0094] Using the method of Example 1 to predict the methylation status of the sample to be tested

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Abstract

The invention relates to the field of molecular biology detection, in particular to a primer, a kit, a model and a construction method for the methylation of cervical cancer-related genes. The methylation primer is designed, and cervical cell DNA is extracted, and the cervical cell DNA is methylated. Methylation treatment, using a PCR amplification kit to perform the first PCR amplification and purification on the cervical cell DNA after methylation treatment, and performing the second PCR amplification and purification on the treated product to obtain a library. The library was subjected to high-pass sequencing on the next-generation sequencing platform for bioinformatics analysis. Based on cervical cancer samples and normal samples, a methylation score was calculated for each PCR amplification region as an independent variable, and the correlation between cervical cancer and cervical cancer was constructed by logistic regression. Gene methylation evaluation model; the present invention builds a database through multiple PCR methylation, and obtains the methylation status of the sample through high-throughput sequencing. The sequencing depth is high, the time is short, and the cost is low. According to the assessed risk, it is decided whether to treat or observe , can effectively reduce overtreatment.

Description

technical field [0001] The invention relates to the field of molecular biology detection, in particular to a primer, a kit, a model and a construction method for the methylation of cervical cancer-related genes. Background technique [0002] Current cervical cancer screening includes cervical cytology testing and human papillomavirus (HPV) testing. Cytology tests include Pap smears and TCT liquid-based cell pellets. Compared with Pap smear, TCT is easier to find abnormal cells and has higher sensitivity, but the price is relatively high, and the diagnostic level is greatly affected by the subjective factors of doctors. Compared with cytological detection, HPV DNA detection can perform typing and quantitative detection of high-risk and low-risk types, but more than 80% of women will be infected with HPV in their lifetime, and more than 90% of infected people will automatically clear the virus within two years , HPV DNA testing produces a large number of false positives (low...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6806C12Q1/686C40B50/06G16B5/00G16B25/20G16B30/10
CPCC12Q1/6886C12Q1/686C12Q1/6806C40B50/06G16B5/00G16B25/20G16B30/10C12Q2600/154C12Q2537/143
Inventor 伍建姬晓雯王海丽
Owner 北京迈基诺基因科技股份有限公司
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