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Reagent for liver cancer diagnosis, kit, detection method and application

A kit and liver cancer technology, applied in the field of molecular biology, to achieve the effect of convenient material collection, good stability and low cost

Active Publication Date: 2018-01-26
上海数熵维度医学信息科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In short, when unmethylated, genes are expressed normally, but when methylated, gene expression is instead suppressed through the process of certain regulatory systems

Method used

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  • Reagent for liver cancer diagnosis, kit, detection method and application
  • Reagent for liver cancer diagnosis, kit, detection method and application
  • Reagent for liver cancer diagnosis, kit, detection method and application

Examples

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Embodiment example

[0040] As a more preferred embodiment of the present invention, the method includes the following steps:

[0041] (1) Extract and purify genomic DNA from clinical samples;

[0042] (2) using chemical treatment or enzymatic treatment of the genomic DNA to distinguish methylated cytosine from unmethylated cytosine;

[0043] (3) amplifying the region containing methylated cytosine in the NEBL gene, FAM55C gene, GALNT3 gene and DSE gene of the genomic DNA by PCR;

[0044] (4) Determining the content of liver cancer-specific methylated cytosine in regions containing CpG sequences in the NEBL gene, FAM55C gene, GALNT3 gene, and DSE gene in the genomic DNA.

[0045] Wherein the content of liver cancer-specific methylated cytosine in the NEBL gene, FAM55C gene, GALNT3 gene, and DSE gene in the clinical sample is the following judgment indications of the clinical sample: suffering from liver cancer; increased risk of liver cancer; recurrence of liver cancer risk.

[0046] In some mo...

Embodiment 1

[0059] 1. Screening of liver cancer-specific cytosine methylation regions in NEBL gene, FAM55C gene, GALNT3 gene and DSE gene

[0060] The screening method is as follows:

[0061] (a) Download the methylation microarray data of TCGA hepatocellular carcinoma, including the methylation profiles of 375 liver cancer tissues and 50 paracancerous tissues. Download the RNA sequencing data of TCGA hepatocellular carcinoma, including gene expression profiles of 369 liver cancer tissues and 50 paracancerous tissues. Download the methylation array data GSE69270 from GEO, including the methylation profiles of 184 non-tumor patients' blood. Download methylation microarray data of other tumor types (BLAC, BRCA, COAD, GBM, HNSC, KIRC, LUAD, LUSC, READ and UCEC) from TCGA.

[0062] (b) Analyze the methylation profile of 50 cancer-adjacent cancer pairings, compare the methylation levels of each site in cancer and para-cancer by paired test, and convert the P value into FDR value by multiple ...

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Abstract

The invention discloses a reagent for liver cancer diagnosis, a kit, a detection method and application. The reagent for liver cancer diagnosis comprises PCR (Polymerase Chain Reaction) primer pairs which aim to at least two of an NEBL gene, an FAM55C gene, a GALNT3 gene and a DSE gene of genome DNA and comprise methylated cytosine. By adopting the reagent, six areas with methylated cytosine on four genes are subjected to conjoint analysis as biomarkers for judging liver cancer pathogenesis and reoccurrence risk, the accuracy of the biomarker is far greater than that of detection of a single biomarker, and false positive is reduced. In addition, the reagent has the advantages of being convenient in material taking, low in cost, high in sensitivity, good in stability and the like, the pathogenesis and reoccurrence risk of cancer liver can be precisely detected in time, great clinical application values can be achieved for rapid diagnosis on early-stage cancer liver, and further researchon rapid diagnosis kits for patients suffering from early-stage liver cancer is technically ensured.

Description

technical field [0001] The present invention relates to the technical field of molecular biology, more specifically, to the technical field of DNA, in particular to a method of using the degree of cytosine methylation in NEBL gene, FAM55C gene, GALNT3 gene and DSE gene to jointly diagnose liver cancer and liver cancer Reagents, kits, detection methods and their uses for risk of incidence and recurrence risk of liver cancer. Background technique [0002] Liver cancer is a tumor that seriously threatens the health and life safety of the people in our country. In my country, the incidence of liver cancer ranks second among malignant tumors. Liver cancer has a high degree of malignancy, because there are often no symptoms or no obvious symptoms at the beginning of the onset, and the tumor progresses rapidly. At the time of diagnosis, about 70-80% of the patients cannot receive surgical treatment, the treatment effect is very poor, and the survival period is very short. . Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
Inventor 丁国徽李虹魏冬凯李光闻彬
Owner 上海数熵维度医学信息科技有限公司
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