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Marker and probe composition for screening bladder cancer and application of marker and probe composition

A technology of markers and compositions, applied in the field of probe compositions and markers for bladder cancer screening, can solve the problems of lack of non-invasive diagnostic methods and achieve high sensitivity and accuracy

Pending Publication Date: 2022-05-27
BIOCHAIN BEIJING SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Early detection is the key to improving patient survival and cure rates, but accurate non-invasive diagnostic methods are currently lacking

Method used

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  • Marker and probe composition for screening bladder cancer and application of marker and probe composition
  • Marker and probe composition for screening bladder cancer and application of marker and probe composition
  • Marker and probe composition for screening bladder cancer and application of marker and probe composition

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0117] The preparation of the chip is mainly based on a glass sheet or a silicon sheet as a carrier, and oligonucleotide fragments or cDNAs are arranged on the carrier in sequence as probes by means of in situ synthesis and micro-array.

[0118] The chip described in this application is based on the signal detection of DNA sequence hybridization after bisulfite treatment. Bisulfite treatment is to change unmethylated cytosine into uracil, while methylated cytosine remains unchanged. Change, then convert uracil to thymine, and finally carry out chip hybridization; finally judge the type of base added according to the fluorescent color, and then determine whether the site is methylated.

[0119] The present application provides a method for bladder cancer screening, which includes:

[0120] detect the methylation level of the marker, and

[0121] The risk of the subject suffering from bladder cancer is judged based on the methylation level, and the marker is selected from one o...

Embodiment 1

[0125] Example 1. Screening markers

[0126] 1) Sample collection: Download the 450k methylation microarray cancer tissue data in TCGA, involving a total of 7,769 cancer tissue samples of 26 types of tumors, including adrenocortical carcinoma (80), bladder urothelial carcinoma (409), acute myeloid Leukemia (140), brain low-grade glioma (654), breast cancer (740), cervical cancer (286), colorectal cancer (348), esophageal cancer (183), uveal melanoma (80), head and neck squamous Stem cell carcinoma (527), bladder tissue carcinoma (660), liver cancer (377), lung adenocarcinoma (425), lung squamous cell carcinoma (372), diffuse large B-cell lymphoma (29), ovarian serous cystadenocarcinoma (10), pancreatic cancer (184), mesothelioma (116), prostate cancer (488), skin melanoma (104), sarcoma (117), gastric cancer (397), testicular cancer (134), thymus cancer (94) ), thyroid cancer (506), endometrial cancer (309). For healthy people, Boercheng collected the plasma of 38 healthy pe...

Embodiment 2

[0139] Example 2. Sample preparation, library building and verification methods

[0140] A marker that can distinguish bladder cancer from healthy people obtained in Example 1 was verified in a plasma sample, and the specific detection method was as follows:

[0141] 1.1.cfDNA extraction and purification

[0142] 1.1.1. Plasma sample preparation:

[0143] Centrifuge the blood sample at 2000 g for 10 min at 4°C and transfer the plasma to a new centrifuge tube. Centrifuge the plasma sample at 16,000g at 4°C for 10 minutes, and proceed to the next step according to the type of collection tube used. The type of collection tube used in this experiment is other.

[0144] Table 1

[0145]

[0146] 1.1.2. Lysis and conjugation

[0147] 1.1.2.1. Prepare the binding solution / bead mixture according to the table below and mix thoroughly.

[0148] Table 2

[0149]

[0150] Add an appropriate volume of plasma sample.

[0151] 1.1.2.2. Thoroughly mix the plasma sample and bindin...

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Abstract

The invention discloses a marker for bladder cancer screening, a probe composition and application thereof. The marker is selected from any one of three markers. According to the present invention, with the application of the marker, the methylation state of the gene can be sensitively and specifically detected so as to be used for the detection of the peripheral blood free DNA, and the composition is used for the screening of the asymptomatic population in the non-invasive manner so as to reduce the harm caused by the invasive detection; the composition has higher sensitivity and accuracy, and can realize real-time monitoring.

Description

technical field [0001] The present application relates to the field of biotechnology, in particular to a marker for bladder cancer screening, a probe composition and applications thereof. Background technique [0002] The bladder is an important part of the human urinary system. Bladder cancer is one of the malignant tumors with the highest incidence in the male genitourinary system. According to the 2020 global cancer report released by WHO, its morbidity and mortality are at the forefront. Among them, its morbidity and mortality rate ranked 6th (4.4%) and 10th (2.9%), respectively. [0003] Because of the obvious symptoms, the vast majority of bladder cancers can be diagnosed early, and most of the diagnosed bladder cancers are superficial non-invasive bladder cancers, a large part of which are stage 0 and stage I, and only need surgery-based treatment can achieve very good results. Generally speaking, the 5-year survival rate of stage 0 bladder cancer is as high as 98%,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11C12N15/12C40B40/06
CPCC12Q1/6886C40B40/06C12Q2600/154
Inventor 连明韩晓亮王小奇刘栓平魏闯张聪杨亚东
Owner BIOCHAIN BEIJING SCI & TECH
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