One group of methylated genes and detection method thereof

A detection method and methylation technology, applied in the field of medical detection, can solve the problems of low early diagnosis rate, invasiveness, and large damage, and achieve the effect of high sensitivity and specificity, and simple operation.

Inactive Publication Date: 2017-06-20
KUNMING UNIV OF SCI & TECH
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Problems solved by technology

[0006] The purpose of the present invention is to provide a group of methylated genes and its detection method, aiming at solving the difficulty in the early diagnosis of lung cancer, the early diagnosis rate is low, and it is difficult to find by imaging methods (such as conventional CT examination) The results of the traditional bronchial brushing method and sputum exfoliation cytology method are not satisfactory

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  • One group of methylated genes and detection method thereof

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[0079] Example 1. The methylation detection of p16, DLEC1, CDH1, DAPK and RUNX3 genes in all the above plasma samples.

[0080] In this embodiment, the nMSP method is used to detect the methylation status of the above five genes, and the specific steps are as follows:

[0081] Step 1: Separation and storage of plasma samples:

[0082] Among all the above blood samples, about 20ml of peripheral blood is taken from each patient or normal person, and immediately stored in a refrigerator at 4°C for a short period of time (about 1h) to facilitate subsequent plasma separation.

[0083] Put two anticoagulation tubes each containing about 10ml of peripheral blood samples in a low-speed centrifuge, and then centrifuge at 2500 rpm at 4°C for 15 minutes to separate the samples into three layers (ie, plasma, white blood cells, and red blood cells). Use a pipette to carefully and gently aspirate the uppermost plasma in the anticoagulation tube (aspirate 2 / 3) and place it in a new 15ml centrifuge t...

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Abstract

The invention discloses one group of methylated genes and a detection method thereof. The group of methylated genes comprises genes p16, DLEC1, CDH1, DAPK and RUNX3; DNA (Deoxyribonucleic Acid) gene sequences are SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5 respectively. In a free DNA sample of plasma of a patient suffering from a lung cancer, specific CpG (Cytosine-Phosphate-Guanine) loci of the genes generate hypermethylation. Those genes provided by the invention are biomarkers for the lung cancer, can be used as a foundation for designing a diagnostic reagent for the lung caner, are applicable to the early auxiliary detection of the lung cancer in a hospital, and can be used for carrying out early risk evaluation on a lung-cancer high risk group.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a group of methylated genes and a detection method thereof. Background technique [0002] Lung cancer is a common malignant tumor. In recent decades, its morbidity and mortality have been on the rise in the world, and it often ranks first among malignant tumors. Many studies have shown that the occurrence and development of lung cancer is a multi-gene, multi-factor and multi-stage complex process, which roughly involves two major mechanisms, one is epigenetics level, and the other is genetics level. Among them, epigenetics plays a pivotal role in the occurrence and development of lung cancer, and DNA methylation is the most important modification method of epigenetics. [0003] DNA methylation is an endogenous modification process in eukaryotic organisms. It generally refers to the conversion of S-adenosylmethionine (SAM-CH3) by organisms due to the catalys...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q1/6848C12Q1/6858C12Q2600/154
Inventor 孔祥阳武超群黄新伟付玉郭丽琼
Owner KUNMING UNIV OF SCI & TECH
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