Unlock instant, AI-driven research and patent intelligence for your innovation.

A reagent combination for detecting liver cancer, kit and application thereof

A technology of detection reagents and kits, which is applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc.

Active Publication Date: 2022-06-14
SANSURE BIOTECH INC
View PDF11 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This project has been certified by the FDA as a breakthrough medical device, but its clinical research is mainly based on European and American populations and has the defect of low sensitivity; the patent CN107164508A detects liver cancer by detecting the content of 5-hydroxymethylcytosine in 9 genes, which has a relatively good performance The detection performance is high, with a sensitivity of 90% and a specificity of 91.3%, but the included samples do not include liver cirrhosis, hepatitis and other high-risk groups of liver cancer, so there may be a deficiency of low specificity

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A reagent combination for detecting liver cancer, kit and application thereof
  • A reagent combination for detecting liver cancer, kit and application thereof
  • A reagent combination for detecting liver cancer, kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1, the screening of methylation gene

[0067] The present invention collected 785 cases of cancer tissues, 461 cases of paracancerous tissues or normal controls from the TCGA data set (https: / / tcga.xenahubs.net) of the UCSC Xena website and the GEO database of the National Center for Biotechnology Information (NCBI) Tissue and 656 healthy whole blood methylation data. Differential analysis was performed with liver cancer and control data, and the physical location and genetic information annotation of the differential sites were performed. In order to ensure that the screened fragments have a consistent methylation level, the screening of methylated gene fragments must meet the following requirements at the same time: 1) It is required that the selected gene fragments have no less than 2 adjacent sites with consistent methylation levels; 2) Perform differential analysis with liver cancer and paracancerous tissues or normal control tissues, and select liver ca...

Embodiment 2

[0068] Example 2. Detection of gene methylation level in clinical samples

[0069] Collect 10ml of each sample of peripheral blood for detection and analysis of the methylation level of DNA methylation markers in the sample. The experimental procedure is as follows:

[0070] 1. Sample preparation

[0071] The sample preparation of the present invention is by MagMAX TM Cell-Free DNA Isolation Kit extract 4ml plasma, 45μL eluent elution. The extracted free nucleic acid needs to meet the following quality control conditions: the total amount of extracted nucleic acid is greater than 20ng.

[0072] 2. Library preparation

[0073] In the present invention, all free nucleic acids qualified for quality control are treated with bisulfite using EZ DNA Methylation-Lightning TM Kit (Zymo Research, Irvine, CA, USA). Subsequently, the sample DNA treated with bisulfite was used to construct a pre-library using the single-strand library construction method. After the pre-library passed ...

Embodiment 3

[0081] Example 3, the reagent combination of the present invention detects the methylation level of clinical samples

[0082] Samples of 63 patients with primary liver cancer, 25 patients with liver cirrhosis, 15 patients with hepatitis and 7 healthy people were collected, and the methylation level of the OTX1 gene in the samples was detected and analyzed according to the method described in Example 2 to verify It examines the effect of liver cancer. The diagnosis of hepatocellular carcinoma, liver cirrhosis, hepatitis and healthy people were all based on the final hospital pathological diagnosis. The test results are shown in Table 1, figure 1 and figure 2 shown.

[0083] Table 1 Prediction performance of OTX1 gene methylation sites in the Logistics liver cancer classification model

[0084]

[0085] It can be seen from Table 1 that each methylation site of the OTX1 gene in tissues can predict liver cancer with a specificity of at least 0.846, a sensitivity of 0.860, ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Login to View More

Abstract

The present invention provides a reagent combination for detecting liver cancer, the reagent combination includes detection reagents for detecting the methylation level of at least one of the following methylation sites of the OTX1 gene: cg23229261, cg10122865. At the same time, the present invention also provides the use of the reagent combination and a kit comprising the reagent combination. Using the reagent combination of the present invention, liver cancer can be predicted with a specificity of at least 0.846 and a sensitivity of 0.860 and an area under the curve of 0.914 in tissue samples, and can be predicted with a specificity of at least 0.8298 and a sensitivity of 0.778 in plasma cell-free DNA samples. The sensitivity and the area under the curve of 0.8823 are used to predict liver cancer, and the reagent combination of the present invention can detect liver cancer with high sensitivity and good specificity in clinical practice with less markers, saving both cost and time; clinically , can be detected sensitively and specifically in the early stage of liver malignant transformation.

Description

technical field [0001] The invention belongs to the field of molecular biology detection, specifically to the field of liver cancer detection, and more specifically to the detection of the methylation level of liver cancer gene markers. Background technique [0002] Liver cancer is one of the most common and deadly diseases in the world, and it is a malignant tumor with great harm. The onset of liver cancer is insidious, and there are no specific symptoms in the early stage. Most liver cancer patients are already in the middle and late stages when they see a doctor. The surgical cure rate is low and the survival period is short. [0003] At present, serum alpha-fetoprotein (alpha fetal protein, AFP) and ultrasonography are commonly used and important methods for clinical diagnosis of liver cancer, but their sensitivity and specificity are not ideal. The sensitivity of AFP screening can only reach 40-60%, and the AFP level of many patients with early liver cancer is always m...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 陈明郭鑫武洪梅刘让蛟戴立忠
Owner SANSURE BIOTECH INC