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Gene markers for detecting liver cancer and uses thereof

A gene marker, liver cancer technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as loss of treatment period, elevated serum AFP, imaging effects, etc.

Active Publication Date: 2021-03-09
北京玄鸟飞讯科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, imaging is easily affected by operator experience, and depends on equipment, which is expensive, especially in the case of limited medical resources, its accuracy is difficult to guarantee, and it is difficult to be widely and routinely applied
The detection sensitivity of alpha-fetoprotein is difficult to exceed 60%, but the sensitivity and specificity of AFP to early liver cancer are not high. For example, in some patients with chronic liver diseases other than liver cancer, such as many patients with chronic hepatitis and liver cirrhosis, serum AFP also increases
This makes most patients diagnosed as advanced liver cancer, losing the best treatment period

Method used

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  • Gene markers for detecting liver cancer and uses thereof
  • Gene markers for detecting liver cancer and uses thereof
  • Gene markers for detecting liver cancer and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. Screening of Liver Cancer Gene Markers

[0028] (1) Extract plasma DNA:

[0029] 10ng plasma DNA was extracted from 20 liver cancer samples with hepatitis B and 20 hepatitis B samples respectively. This step can be carried out using any methods and reagents suitable for extracting plasma DNA well known to those skilled in the art.

[0030] (2) End-fill the plasma DNA, overhang A and connect with the sequencing adapter:

[0031] Prepare a reaction mixture containing 50uL plasma DNA, 7uL End Repair&A-Tailing Buffer and 3uL End Repair&A-Tailing Enzyme mix according to the Kapa Hyper Perp Kit instructions (total volume is 60uL), incubate at 20°C for 30 minutes, then incubate at 65°C for 30 minutes minute. Prepare the following ligation reaction mixture in a 1.5mL low adsorption EP tube: 5uL Nuclease free water, 30uL Ligation Buffer and 10 uL DNA Ligase. Add 5uL of sequencing adapters to 45uL ligation reaction mixture, mix, heat at 20°C for 20 minutes, and the...

Embodiment 2

[0051] Example 2. Effectiveness of Liver Cancer Gene Markers

[0052] This example verifies the effectiveness of the liver cancer gene markers of the present invention for detecting liver cancer.

[0053] Measure the 5-hmC content of 11 liver cancer gene markers according to the present invention in 110 samples (60 routine liver cancer samples with hepatitis B and 50 routine hepatitis B samples) according to the method of embodiment 1, and determine the weighting of each genetic marker coefficient.

[0054] The normalized 5-hmC content of each gene marker is multiplied by the corresponding weighting coefficient to obtain the predictor t of the gene marker, and then the predictor t of each gene marker is added to obtain the total predictor T, and then The total predictor T was Logistically transformed according to the following formula to obtain the score P:

[0055]

[0056] If P>0.5, the subject sample has liver cancer; if P≤0.5, the subject sample does not have liver ca...

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Abstract

The invention relates to a gene marker for detecting liver cancer and its application for hepatitis B patients. The present invention also relates to a method for detecting liver cancer using the gene marker.

Description

technical field [0001] The present invention relates to the field of clinical molecular diagnosis of liver cancer. Specifically, the present invention relates to a method and a kit for detecting the presence of liver cancer by detecting the content of 5-hydroxymethylcytosine in liver cancer gene markers through high-throughput sequencing. Background technique [0002] Liver cancer is one of the most common malignancies worldwide. According to the statistics of the World Health Organization in 2008, there are 748,300 new cases and 695,900 deaths in the world every year, of which more than 50% occur in China. According to statistics, more than 400,000 cases of liver cancer are newly diagnosed every year in my country, and more than 85% of liver cancer patients have been infected with hepatitis B virus. China's existing chronic hepatitis B infection accounts for a quarter of the world's total, and liver cancer cases account for half of the world's total. Chronic hepatitis inf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/154
Inventor 陆星宇宋艳群彭莱
Owner 北京玄鸟飞讯科技有限公司
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