Methylation gene combination, primer and probe combination and kit for cervical cancer methylation detection and using method of kit

A gene combination and methylation technology, applied in the field of medical gene detection, can solve problems such as transient and excessive treatment, and achieve the effect of reducing protein expression and reducing randomness

Pending Publication Date: 2019-11-15
韩林志
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Because it only detects the presence or absence of HPV virus, and the presence of HPV virus is mostly transient, which m

Method used

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  • Methylation gene combination, primer and probe combination and kit for cervical cancer methylation detection and using method of kit
  • Methylation gene combination, primer and probe combination and kit for cervical cancer methylation detection and using method of kit
  • Methylation gene combination, primer and probe combination and kit for cervical cancer methylation detection and using method of kit

Examples

Experimental program
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Embodiment 1

[0056] Example 1 Application of the kit of the present invention in the diagnosis of human cervical cancer

[0057] The nuclease-free water, 10×PCR Buffer, HS Taq enzyme, and dNTPs used in the present invention were purchased from Yubao Biology (Dalian) Co., Ltd.;

[0058] Positive control (cervical cancer cell line (CaSki)), negative control (cervical cancer cell line (C33A)).

[0059] The DNA extraction kit is: "Nucleic Acid Extraction and Purification Reagent" from Hunan Baibo Gene Technology Co., Ltd.;

[0060] The transformation kit is, EZ DNA Methylation TM Kits.

[0061] The biological materials used in the present invention are all from Xiangya Medical Laboratory.

[0062] method:

[0063] 1. Biological samples:

[0064]From January 2017 to June 2017, 33 cases of C1N1 cervical exfoliated cells, 30 cases of C1N2 cervical exfoliated cells, 30 cervical exfoliated cells of C1N3+ cervical cancer patients, and 90 cases of cervical exfoliated cells of normal people in th...

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Abstract

The invention provides a methylation gene combination, primer and probe combination and kit for cervical cancer methylation detection and a using method of the kit, and belongs to the technical fieldof medical gene detection. Based on a real-time fluorescent PCR technology, specific methylation detection primers and probes in the promoter regions of FAM19A4, has-miR-124-2 and ZNF671 genes are designed and prepared into a PCR reaction solution to amplify a sample DNA after vulcanization, the quality of the sample is controlled by means of an internal reference gene beta-actin, and whether or not the sample is methylated is determined. Compared with the conventional method, the method has the advantages of fast speed, high flux and good sensitivity and specificity, and plays an important role in the early diagnosis and prognosis of cervical cancer.

Description

technical field [0001] The invention relates to the technical field of medical gene detection, in particular to a methylation gene combination, a primer and probe combination, a kit and a use method thereof for cervical cancer methylation detection. Background technique [0002] Cervical cancer has always been an important malignant tumor that plagues women. Normal cervical cells develop from precancerous changes to cervical cancer, which may take decades. During the whole process, some HPV virus infections can be repaired by the autoimmune system and return to normal automatically, while other precancerous lesions that are more closely related to cervical cancer need to be intervened in time to prevent the precancerous lesions from developing into cervical cancer . [0003] At present, the main screening methods used clinically in my country are cytological detection methods, including Pap smear and liquid-based cytology (TCT). However, the detection process of cytologic...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/154C12Q2600/178C12Q2523/125C12Q2531/113C12Q2563/107
Inventor 韩林志肖芳李翔李书
Owner 韩林志
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