Novel multi-target combined marker for early detection of liver cancer and application thereof

A marker, a technology for liver cancer, applied in measurement devices, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problem of inability to distinguish liver cancer and hepatitis patients, low sensitivity and specificity, and low liver cancer AUC value, etc. problem, to achieve the effect of consistent test results, high test results, and improved effect

Active Publication Date: 2022-06-07
杭州翱锐基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the AUC value of the existing methylation site detection of early liver cancer is generally low, the sensitivity and specificity are not high, and it cannot distinguish liver cancer and hepatitis patients well, and further examinations are still needed to confirm the diagnosis

Method used

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  • Novel multi-target combined marker for early detection of liver cancer and application thereof
  • Novel multi-target combined marker for early detection of liver cancer and application thereof
  • Novel multi-target combined marker for early detection of liver cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1 Screening of gene methylation sites

[0090] In this example, high-depth (40×-60×) whole-genome methylation sequencing ( WGBS), selected the differentially methylated regions of 300 genes in the high-depth WGBS data, including both hypermethylated and hypomethylated regions, and efficiently differentiated liver cancer and hepatitis B patients ( figure 1 ). At the same time, a large amount of public data (>1,000 450K microarray data of liver cancer and adjacent cancer cases) was synthesized, and the target region was reduced to 200 genes and 6000 CpG regions.

[0091] In order to avoid the sampling bias of the 200 genes and 6000 CpG regions screened in the previous stage, we further designed a methylation capture panel for these 200 genes, and conducted more than 500 cases of different stages of liver cancer and high-risk groups of liver cancer (hepatitis B and liver cirrhosis). ) cfDNA samples for ultra-high depth (>10,000X) next-generation sequencing to bui...

Embodiment 2

[0093] Example 2 Screening of early liver cancer using 3 methylation sites (or binding protein markers)

[0094] In this example, 3 of the 33 gene methylation sites screened in Example 1 were selected for the detection of early liver cancer. Two methods were used for detection: first, using three methylation sites to detect early-stage liver cancer; second, using a combination of three methylation sites and protein markers to detect early-stage liver cancer.

[0095] 1. Using 3 methylation sites to detect early liver cancer

[0096] The specific method for detecting early-stage liver cancer using three methylation sites includes the following steps:

[0097] In the first step, the serum and plasma of the blood samples were separated, and the magnetic bead method was used to extract the reagents to extract the plasma-free DNA of the biological samples to be tested. Among them, 20 were liver cancer patients and 20 were hepatitis patients.

[0098] In the second step, the methy...

Embodiment 3

[0131] Example 3 Screening of early-stage liver cancer using 6 methylation sites (or binding protein markers)

[0132] In this example, 6 of the 33 gene methylation sites screened in Example 1 were selected for the detection of early liver cancer. Two methods were used for detection: first, using 6 methylation sites to detect early liver cancer; second, using the combination of 6 methylation sites and protein markers to detect early liver cancer.

[0133] 1. Using 6 methylation sites to detect early liver cancer

[0134] The present embodiment selects the following groups of combinations comprising 6 methylation sites to detect respectively:

[0135] 4. Seq ID NO.1, Seq ID NO.12, Seq ID NO.13, Seq ID NO.25, Seq ID NO.30, SeqID NO.31

[0136] 5. Seq ID NO.1, Seq ID NO.12, Seq ID NO.23, Seq ID NO.25, Seq ID NO.27, SeqID NO.28

[0137] 6. Seq ID NO.1, Seq ID NO.8, Seq ID NO.12, Seq ID NO.18, Seq ID NO.25, SeqID NO.26

[0138] 7. Seq ID NO.1, Seq ID NO.2, Seq ID NO.3, Seq ID N...

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Abstract

The invention provides a novel multi-target combined marker for early detection of liver cancer and application thereof, a series of novel methylation sites capable of efficiently distinguishing liver cancer and hepatitis patients are found from high-depth whole genome methylation (WGBS) data of liver cancer tissues, para-carcinoma tissues and hepatitis patient samples, and the novel combined marker can be used for efficiently detecting early liver cancer; the novel gene methylation site is combined with other liver cancer detection markers (such as AFP, AFP-L3 and DCP), the AUC value can be further increased, and the sensitivity and specificity of early liver cancer screening are improved.

Description

[0001] This application claims the priority of the Chinese earlier application, application number: 202111439557.1, and the filing date is November 30, 2021; all contents thereof are regarded as a part of the present invention. technical field [0002] The invention relates to the field of early cancer screening, in particular to a novel multi-target combination marker for early detection of liver cancer and its application. Background technique [0003] Liver cancer is the most common tumor and the leading cause of cancer death in men under the age of 60. In recent years, my country's liver cancer diagnosis and treatment technology has made great progress, but the overall 5-year net survival rate of patients after age standardization has only increased from 11.7% in 2000-2004 to 14.1% in 2010-2014, and no significant improvement has been seen. . However, the 5-year overall survival rate of Barcelona liver cancer clinical stage (BCLC) 0 or A stage liver cancer patients who r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6827C12N15/11G01N33/574G01N33/573
CPCC12Q1/6886C12Q1/6827G01N33/57438G01N33/57476G01N33/573C12Q2600/154C12Q2531/113C12Q2563/107Y02A50/30
Inventor 张琼朱友杰徐博
Owner 杭州翱锐基因科技有限公司
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