Gene marker for detecting liver cancer and purpose thereof
A technology of gene markers and liver cancer, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of lost treatment period, difficulty in ensuring accuracy, difficulty in widespread and routine application, etc.
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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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