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A nucleic acid mass spectrometry detection method for early screening of liver cancer susceptibility genes

A technology for gene detection and liver cancer, applied in biochemical equipment and methods, microbe measurement/inspection, etc., can solve problems affecting the sensitivity of liver cancer

Active Publication Date: 2021-02-19
武汉赛云博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some HCC patients maintain a low level of serum AFP (<20ng / ml) for a long time, and B-ultrasound has certain limitations in detecting and further judging the characteristics of nodules, which affects the sensitivity of early diagnosis of liver cancer
At present, there is no very effective early detection method in clinical practice, so it is particularly important to establish a highly sensitive, economical and simple molecular screening method

Method used

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  • A nucleic acid mass spectrometry detection method for early screening of liver cancer susceptibility genes
  • A nucleic acid mass spectrometry detection method for early screening of liver cancer susceptibility genes
  • A nucleic acid mass spectrometry detection method for early screening of liver cancer susceptibility genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0140] Example 1: Feasibility analysis of SNPs of human liver cancer susceptibility-related genes and screening of gene mutation sites related to liver cancer

[0141] The inventors of the present invention searched NCBI domestic and foreign genome-wide association study (genome-wide association study, GWAS) research literature and related patents (patent literature 5 and non-patent literature 11, etc., including but not limited to This), while searching the GWAS Catalog database and cancer-related databases such as COSMIC, CIViC, ICGC, and MYCANCER GENOME, screened out more than 1,000 positive samples from the Asian population and more than 500 positive samples from the Chinese population in the GWAS study. The inventors screened and evaluated the liver cancer-related sites verified in the research, and selected 4 liver cancer hotspot mutations and 23 single nucleotide polymorphism sites that are significantly related to the risk of liver cancer in Asian populations , and are...

Embodiment 2

[0160] Embodiment 2: Detection of liver cancer cell lines by the technology of the present invention

[0161] 1. Research object

[0162] It is feasible to select six liver cancer cell lines (all of which can be purchased from ATCC) with known mutation sites SNU-475, C3A, PLC / PRF / 5, SK-HEP-1, SNU-387, and SUN-449 for the detection method For sex analysis, normal human genome DNA (gDNA) (extracted from the oral mucosa tissue of normal people in Wuhan General Hospital of Guangzhou Military Region) was set up as a negative control, and water (H 2 0) as a blank control.

[0163] 2. Experimental steps

[0164] (1) Extract DNA to obtain a DNA extract;

[0165] (2) As shown in Table 1, respectively mix 19 pairs of amplification primers of P1 group, 7 pairs of amplification primers of P2 group and 1 pair of amplification primers of P3 group, and add the mixed specific amplification primers to DNA Extract the solution, and then carry out PCR amplification reaction in the amplificat...

Embodiment 3

[0190] Example 3: Detection of SNPs and mutation sites of genes related to early diagnosis of liver cancer in peripheral blood samples

[0191] 1. Research object

[0192] In this study, peripheral venous blood samples from Wuhan General Hospital of Guangzhou Military Command were used for genetic testing of samples, and the number of cases was 30. The control group was a sample of healthy people from local health checkups.

[0193] 2. Experimental steps

[0194] The specific steps of applying the technology of the present invention to detect the SNPs and mutation sites of genes related to the early diagnosis of liver cancer in peripheral blood samples are as follows:

[0195] (1) Use the rapid extraction kit to extract cfDNA:

[0196] (a) Take 200 μl of collected peripheral blood and put it into a 1.5ml centrifuge tube. Add 200 μl of binding solution CB, shake it upside down vigorously immediately, and mix well, then add 20ul proteinase K (20mg / ml) solution, shake it upsi...

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Abstract

The invention relates to the field of gene detection, in particular to a nucleic acid mass spectrometry detection method for early screening of liver cancer susceptibility genes. The present invention considers the difference of liver cancer gene spectrum between the Chinese population and the European and American population, screens out the combination of SNPs of genes related to Chinese liver cancer susceptibility and gene mutation sites related to liver cancer, and uses nucleic acid mass spectrometer to analyze the genetics related to liver cancer. Extensive (high-throughput detection sites, high-throughput detection samples) screening and inspection are performed on the logo. The method of the present invention has a high detection success rate, good technical reproducibility, and high cost performance, and can realize the detection of multiple genes in a single small sample to meet the maximum use of small samples; the method of the present invention is based on MassARRAY nucleic acid mass spectrometry technology, and compared with Sanger Sequencing has the technical advantages of high accuracy and high sensitivity, and the detection results are stable, which improves the positive rate of detection.

Description

technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a nucleic acid mass spectrometry detection method for early screening of liver cancer susceptibility genes. Background technique [0002] Primary liver cancer (Hepatocellular Carcinoma, HCC) is one of the most common and harmful malignant tumors in my country and even in the world. The annual new cases in my country account for about 45% of the world, and it is the third most fatal malignant tumor in the world. Although the curative effect of liver cancer has been greatly improved with the emergence of various new technologies and new treatments and the improvement of diagnosis and treatment, the annual mortality rate of liver cancer in my country is still as high as 20.40 / 100,000, accounting for about 100% of the death rate of liver cancer in the world. 1 / 2. [0003] The occurrence of liver cancer is a complex process involving multiple genes and factors, and is the resu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858C12Q1/6886
Inventor 尚小云刘梦梅刁波
Owner 武汉赛云博生物科技有限公司
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