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DNA methylation biomarker or composition for early liver cancer detection and application of DNA methylation biomarker or composition

A biomarker, early-stage liver cancer technology, applied in recombinant DNA technology, DNA/RNA fragments, combinatorial chemistry, etc., can solve the problems of unsatisfactory liver cancer screening effect, low detection rate and early diagnosis rate, etc. The effect of large-scale promotion and application, improving detection rate and improving prediction accuracy

Active Publication Date: 2022-06-24
北京莱盟君泰国际医疗技术开发有限公司
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  • Claims
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AI Technical Summary

Problems solved by technology

At present, the screening effect of liver cancer is not ideal, and the detection rate and early diagnosis rate are relatively low. How to detect cancer early has become the focus of the medical field. Therefore, a new and convenient early cancer screening method has been developed for individuals. It is very important for society

Method used

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  • DNA methylation biomarker or composition for early liver cancer detection and application of DNA methylation biomarker or composition
  • DNA methylation biomarker or composition for early liver cancer detection and application of DNA methylation biomarker or composition
  • DNA methylation biomarker or composition for early liver cancer detection and application of DNA methylation biomarker or composition

Examples

Experimental program
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Effect test

Embodiment 1

[0052] Example 1 Detection method of plasma cfDNA methylation markers for liver cancer detection

[0053] 1. Plasma cf DNA extraction: The specific operation steps of plasma cf DNA extraction are carried out in accordance with the "Operation Manual of Cell-Free DNA Extraction Kit (Suction Filtration Method)" combined with the vacuum filtration pump: the plasma is cleaved with proteinase K and lysate, and vacuum filtration is used. The pump and DNA-binding column bind the DNA and are washed with washing solution and absolute ethanol, and finally eluted with elution buffer.

[0054] 2. Fragment screening of cfDNA using magnetic beads:

[0055] ①Add 0.75 times the cfDNA product SPRIselect magnetic beads to cfDNA, vortex to mix, and incubate at room temperature for 5 minutes; place on a magnetic stand, and after the solution is clarified, pipette the supernatant into a new 1.5mL centrifuge tube for use ;

[0056] ② Add 1.05 times the SPRIselect magnetic beads of the cfDNA produc...

Embodiment 2

[0154] This example discloses DNA methylation specific markers for detecting liver cancer. Based on 51 liver cancer adjacent samples and 379 liver cancer samples in the TCGA public database, 10 DNA methylation sites with specific and significant methylation differences in liver cancer were mined, including 4 specific methylation sites in liver cancer. Sites with increased methylation and 6 sites with specific decreased methylation in liver cancer. These loci have high AUC values ​​in distinguishing normal liver samples from liver cancer samples, see figure 2 and image 3 . Among them, cg18233405 and cg18842353 performed the best with AUCs of 0.914 and 0.960, respectively. Models constructed using 10 methylation biomarkers can accurately distinguish normal liver samples from early liver cancer samples, see Figure 4 .

Embodiment 3

[0156] In this example, based on 125 plasma samples of benign liver disease and 122 plasma samples of liver cancer, the 10 DNA methylation markers disclosed in the present invention for detection of liver cancer were verified. The AUC values ​​of each methylation marker independently distinguishing liver cancer plasma are shown in Figure 5 and Image 6 .

[0157] The model constructed using the combination of 4 markers with increased methylation and 6 markers with decreased methylation has an AUC value of 0.899 to distinguish benign liver disease plasma from liver cancer plasma, see Figure 7 In A, it shows that all 10 methylation markers have a certain effect on predicting liver cancer, and the model constructed together has the best predictive performance. Since studies have shown that the occurrence of liver cancer is significantly correlated with age and gender, the present invention further combines age, gender and methylation markers to construct an early-stage liver ...

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Abstract

The invention discloses a DNA (deoxyribonucleic acid) methylation biomarker or a DNA methylation biomarker combination for early liver cancer detection, which comprises at least one of the following methylation markers: cg2088488887, cg06437004, cg10483825, cg18233405, cg18766755, cg21769702, cg27134730, cg01578265, cg18842353 and cg06613738. The invention discloses ten specific methylation markers related to occurrence of liver cancer, and the specific methylation markers can be used for remarkably improving the detection rate of early liver cancer. After the methylation marker is further combined with age and gender information, the predicted AUC value of a trained model can reach 0.917 when the model is used for distinguishing benign liver diseases (hepatitis and cirrhosis) and early liver cancer, and the model is suitable for large-scale popularization and application due to noninvasiveness of plasma detection.

Description

technical field [0001] The invention belongs to the technical field of biotechnology, and in particular relates to a DNA methylation biomarker or combination for early liver cancer detection and its application. Background technique [0002] It is well known that early detection and early treatment of cancer can greatly improve the survival rate of patients. The 5-year relative survival rate of liver cancer is only 12.1%, however, if early diagnosis and effective treatment can achieve a relatively good prognosis, especially for small liver cancers within 5 cm, the 5-year survival rate can reach more than 80%. At present, the screening effect of liver cancer is unsatisfactory, and the detection rate and early diagnosis rate are relatively low. How to detect cancer early has become the focus of the medical community. Therefore, a new and convenient early cancer screening method has been developed for individuals. It is very important for society. [0003] DNA methylation i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6806C12Q1/6869C40B50/06C12N15/11
CPCC12Q1/6886C12Q1/6806C12Q1/6869C40B50/06C12Q2600/154C12Q2523/308C12Q2525/173C12Q2525/191C12Q2531/113C12Q2535/122
Inventor 徐建锋李典格张泽跃李清华张素芳
Owner 北京莱盟君泰国际医疗技术开发有限公司
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