Gene marker used for detecting liver cancer and purpose thereof

A gene marker, a technology for liver cancer, applied in the determination/examination of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of low diagnostic accuracy and specificity of small liver cancer, and achieve easy recurrence and metastasis. , easy to operate, high accuracy

Pending Publication Date: 2017-09-15
北京玄鸟飞讯科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, as mentioned above, the accuracy and specificity of these traditional methods for the diagnosis of small liver cancer are not high

Method used

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  • Gene marker used for detecting liver cancer and purpose thereof
  • Gene marker used for detecting liver cancer and purpose thereof
  • Gene marker used for detecting liver cancer and purpose 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 of plasma DNA was extracted from samples from 20 liver cancer patients and 20 normal people. 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 10uL DNA Ligase. Add 5uL of sequencing adapters to 45uL ligation reaction mixture, mix, heat at 20°C for 20 minutes, and then keep at 4°C. The ...

Embodiment 2

[0050] Example 2. Effectiveness of Liver Cancer Gene Markers

[0051] This example verifies the effectiveness of the liver cancer gene markers of the present invention for detecting liver cancer.

[0052]Measure the 5-hmC content of 9 liver cancer gene markers of the present invention in the first batch of 96 samples (50 routine liver cancers and 46 healthy controls) according to the method for embodiment 1, and determine the weighting coefficient of each genetic marker .

[0053] 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:

[0054]

[0055] If P>0.5, the subject sample has liver cancer; if P≤0.5, the subject sample is normal.

[0056] figure 1 The results of dif...

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Abstract

The invention relates to a gene marker used for detecting liver cancer and a purpose thereof. The invention also relates to a method for detecting liver cancer by using the provided 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. Among primary liver cancers, 70%-85% are hepatocellular carcinoma (HCC). At present, the 5-year survival rate of liver cancer is only 3%-5%, because most patients are already in the middle and advanced stages when they see a doctor, and the best time for treatment has been lost. Therefore, early detection, early diagnosis and early treatment are the key to improving th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/158
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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