Methylated primer, kit, model and construction method for cervical cancer related genes

A construction method and methylation technology, applied in biochemical equipment and methods, chemical libraries, combinatorial chemistry, etc., can solve problems such as inaccurate assessment of cervical cancer methylation levels, achieve high sensitivity and specificity, and improve life quality, reducing the effects of overtreatment

Active Publication Date: 2022-01-14
北京迈基诺基因科技股份有限公司
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  • 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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  • Methylated primer, kit, model and construction method for cervical cancer related genes
  • Methylated primer, kit, model and construction method for cervical cancer related genes
  • Methylated primer, kit, model and construction method for cervical cancer related genes

Examples

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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 biological detection, in particular to a methylated primer, a kit, a model and a construction method for cervical cancer related genes. The construction method comprises the following steps: designing a methylated primer; methylating a cervical cell DNA by extracting the cervical cell DNA; performing first PCR amplification and purification on the methylated cervical cell DNA by using a PCR amplification kit; performing second PCR amplification and purification on the treated product to obtain a library; performing high-pass sequencing on the library through a next-generation sequencing platform; performing bioinformatic analysis; calculating each PCR amplification area to obtain a methylation score as an independent variable based on a cervical cancer sample and a normal sample; and constructing a cervical cancer related gene methylation evaluation model by adopting a logistic regression mode. According to the method, the methylation state of the sample is obtained through multiple PCR methylation library establishment and high-throughput sequencing, the sequencing depth is high, the time is short, the cost is low, treatment or observation is determined according to the assessed risk, and excessive treatment can be effectively reduced.

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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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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