Kit and method for predicting hepatocarcinogenesis risk

A kit and risk technology, which are used in kits and fields for predicting the risk of liver cancer, can solve the problems of low sensitivity of blood free nucleic acid detection and narrowing of application scope.

Inactive Publication Date: 2018-05-25
北京旌准医疗科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] However, the above biomarkers are mainly obtained by studying the methylation differences between cancerous tissues and adjacent tissues, and their detection requires more tissue samples, and the detection sensitivity of blood free nucleic acids is not high, so their application scope is greatly reduced , it is difficult to screen large-scale high-risk HCC populations

Method used

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  • Kit and method for predicting hepatocarcinogenesis risk
  • Kit and method for predicting hepatocarcinogenesis risk
  • Kit and method for predicting hepatocarcinogenesis risk

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 Detection of methylation levels of gene combinations by methylation-specific fluorescent PCR method based on bisulfite conversion

[0076] 1. Collection and preparation of blood samples

[0077] 1. Use cfDNA preservation tubes such as Streck Cell-Free DNA BCT blood collection tubes to collect peripheral blood, and the collected volume is 1-10mL.

[0078] 2. Store at 2-8°C or at room temperature within 7 days.

[0079] 3. Take 2mL of peripheral blood collected in the cfDNA preservation tube, and centrifuge to separate the plasma: centrifuge at 1350±150rcf for 12 minutes.

[0080] 2. Extraction of cfDNA and BS conversion

[0081] 1. Use a peripheral blood free DNA extraction kit such as QIAGEN QIAseq cfDNA Extraction Kit to extract cfDNA from the separated plasma and negative and positive quality controls. The extraction of cfDNA in this method adopts the magnetic bead method to absorb, purify, elute and enrich the cfDNA, and can also use the separation colum...

Embodiment 2

[0097] Example 2 Methylation-binding protein enrichment-fluorescent PCR detection method to detect the methylation level of gene combinations

[0098] 1. Collection and preparation of blood samples

[0099] 1. Use cfDNA preservation tubes such as Streck Cell-Free DNA BCT blood collection tubes to collect peripheral blood, and the collected volume is 1-10mL.

[0100] 2. Store at 2-8°C or at room temperature within 7 days.

[0101] 3. Take 2mL of peripheral blood collected in the cfDNA preservation tube, and centrifuge to separate the plasma: centrifuge at 1350±150rcf for 12 minutes.

[0102] 2. Extraction of cfDNA and capture and enrichment of MBD protein

[0103] 1. Use a peripheral blood cell-free DNA extraction kit such as QIAGEN QIAseq cfDNA Extraction Kit to extract cfDNA from the separated plasma. The extraction of cfDNA in this method adopts the magnetic bead method to absorb, purify, elute and enrich the cfDNA, and the method of separation column can also be used.

...

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Abstract

The invention relates to a kit and a method for predicting a hepatocarcinogenesis risk, relating to the technical field of medical in-vitro diagnosis. Particularly, the invention relates to a kit anda method for predicting a hepatocarcinogenesis risk through the CpG island methylation states of single genes or multiple genes in SCT, NFIX, IGF2, ABCG5, GSTO2, SARDH, E2F6, SHANK2, GATA4, OPLAH, NR2F1 and ZFHX3. According to the combined methylation detection of the multiple genes, the sensitivity is greatly improved; the free nucleic acids of saliva, urine, blood and the like are detected, drawing materials from cancer tissue is not required, the detection is noninvasive, and the operation is simple and convenient.

Description

technical field [0001] The present invention relates to the technical field of medical in vitro diagnosis, specifically through the CpG island methylation status of single or multiple genes in SCT, NFIX, IGF2, ABCG5, GSTO2, SARDH, E2F6, SHANK2, GATA4, OPLAH, NR2F1 and ZFHX3 A kit and method for predicting the risk of liver cancer. Background technique [0002] Liver cancer is the fourth most common tumor worldwide. In China, liver cancer is the third most diagnosed tumor after lung cancer and gastric cancer. In 2015, the incidence of liver cancer was 466,000, and the number of deaths due to liver cancer was 422,000. The high morbidity and mortality are due to the fact that the early symptoms of liver cancer are not obvious, and the prediction and early diagnosis of liver cancer are very difficult. Usually, the diagnosis can only be made in the late stage. At this time, the metastasis of tumor cells has often occurred, and the treatment is difficult. It's hard to get better...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686
CPCC12Q1/686C12Q1/6886C12Q2600/118C12Q2600/154C12Q2600/156C12Q2563/107C12Q2537/164
Inventor 林华明刘明坤叶锋
Owner 北京旌准医疗科技有限公司
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