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Reagent for auxiliary diagnosis of liver cancer, and application of reagent in preparation of kit

A technology of auxiliary diagnosis and kit, which is applied in the field of detection of methylation sites, can solve problems such as unsatisfactory sensitivity and specificity, short survival period, hidden onset of liver cancer, etc., and achieve good classification performance

Pending Publication Date: 2020-03-24
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The onset of liver cancer is hidden, and there are no specific symptoms in the early stage. Most patients with liver cancer are already in the middle and late stages when they see a doctor. The cure rate of surgery is low and the survival period is short.
Serum alpha-fetoprotein (alpha fetal protein, AFP), as well as tissue biopsy and ultrasonography are commonly used and important methods for clinical diagnosis of liver cancer, but their sensitivity and specificity are not ideal, and biopsy is invasive

Method used

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  • Reagent for auxiliary diagnosis of liver cancer, and application of reagent in preparation of kit
  • Reagent for auxiliary diagnosis of liver cancer, and application of reagent in preparation of kit
  • Reagent for auxiliary diagnosis of liver cancer, and application of reagent in preparation of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1. Screening of gene methylation sites

[0041] The methylation data and clinical information of 379 cancer tissues and 50 paracancerous tissues were downloaded from the TCGA dataset (https: / / tcga.xenahubs.net) of the UCSC Xena website. The methylation data and clinical information of 355 cases of cancer tissues, 66 cases of paracancerous tissues, 10 cases of normal samples and 9 cases of liver cirrhosis were downloaded from the GEO database of the National Center for Biotechnology Information (NCBI). In addition, 34 pairs of liver cancer tissue samples and their adjacent normal tissue samples from patients with primary liver cancer were collected to detect and analyze the methylation levels of DNA methylation markers in the samples. We label this dataset as LiverNGS, see Table 1.

[0042] Table 1

[0043]

[0044] Differential analysis was performed on the two datasets GSE and LIHC, and shared differentially methylated sites were selected to construct a cl...

Embodiment 2

[0045] Embodiment 2, the analysis method of clinical sample

[0046] Samples of 34 pairs of liver cancer tissues and their adjacent normal tissues collected from patients with primary liver cancer were used to detect and analyze the methylation levels of DNA methylation markers in the samples. In the present invention, relevant liver cancer and adjacent normal tissues were obtained from the hospital, approved by the Medical Research Ethics Committee, and all subjects signed the informed consent. The diagnosis of liver cancer tissue and adjacent normal tissue was based on the final hospital pathological diagnosis. The samples were stored in a -80°C refrigerator until use. The experimental procedure is as follows:

[0047] 1. Sample preparation

[0048] The sample preparation of the present invention is to extract 50 mg of fresh liver cancer tissue and fresh para-cancerous tissue through TIANamp Genomic DNA Kit, and wash it with 100 μL of eluent. The extracted genomic DNA ne...

Embodiment 3

[0087] Example 3, a single methylation site is used to predict liver cancer

[0088] The methylation sites cg22158769 and cg12206199 of LOC375196 were used to predict liver cancer respectively. Based on the classification model of Logistic regression analysis of these two CpG sites, the LiverNGS sequencing data of clinical liver cancer tissue and its adjacent normal tissue samples were used as the validation set. It shows that the classification model has better classification and discrimination ability, and the results are shown in Table 4. It can be seen from the table that a single methylation site of LOC375196 also has better classification performance.

[0089] Table 4

[0090]

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Abstract

The invention belongs to the field of molecular biological detection, particularly belongs to the field of liver cancer detection, provides an application of a reagent in the preparation of a kit forauxiliary diagnosis of liver cancer, and also provides the reagent for auxiliary diagnosis of the liver cancer, and the kit containing the reagent. The reagent can be used for diagnosing liver cancerat least with the sensitivity of 82.35% and the specificity of 100%, and the area under a curve is 0.8720 or above.

Description

technical field [0001] The invention belongs to the field of molecular biology detection, specifically to the field of liver cancer detection, and more specifically to the detection of methylation sites of liver cancer gene markers. Background technique [0002] Liver cancer is one of the most common and deadly diseases in the world, and it is also a malignant tumor with high incidence and great harm in my country. According to Globocan statistics, the number of new cancer patients in the world in 2012 was about 14 million, among which liver cancer patients ranked sixth among common cancer types, and about 782,000 people worldwide suffered from liver cancer, accounting for 5.6% of all new cancer patients . The onset of liver cancer is insidious, and there are no specific symptoms in the early stage. Most liver cancer patients are already in the middle and late stages when they see a doctor. The surgical cure rate is low and the survival period is short. Serum alpha-fetopro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/154
Inventor 陈明郭鑫武洪梅刘让蛟戴立忠
Owner SANSURE BIOTECH INC
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