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A microsatellite biomarker combination, detection kit and use thereof

A technology of biomarkers and detection reagents, applied in the field of genetic screening, to achieve the effect of low price, high specificity and rapid detection

Active Publication Date: 2019-09-06
GUANGZHOU BURNING ROCK DX CO LTD
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, these methods require a sufficient proportion of tumor cells

Method used

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  • A microsatellite biomarker combination, detection kit and use thereof
  • A microsatellite biomarker combination, detection kit and use thereof
  • A microsatellite biomarker combination, detection kit and use thereof

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Embodiment

[0092] 1. Data preparation: based on the next-generation sequencing method for gene combination (panel) detection, the specific steps are as follows:

[0093] The capture steps of tissue samples are as follows: use QIAamp DNA FFPE tissue kit (QIAGEN: 56404) to extract DNA from tumor tissue and adjacent normal tissue respectively. Accurate quantification was performed with the dsDNA HS assay kits (ThermoFisher: Q32854) provided with the Qubit 3.0 Fluorometer. Then, the DNA was physically fragmented into fragments of 180-250 bp using a sonicator Covaris M220 (Covaris: PN500295), followed by end repair, phosphorylation, addition of deoxyadenine to the 3' end, and linker ligation. Then the DNA connected with the amplification adapter was purified with AgencourtAMPure XP paramagnetic beads, and pre-amplified with PCR polymerase, and the amplified purified product was hybridized with the multiple biotin-labeled probe set customized by Agilent (The gene combination (panel) design in...

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Abstract

The present invention relates to biomarker combinations, kits for their detection, and their detection in plasma samples of microsatellite instability (MSI) and non-invasive detection of cancer, preferably colorectal (eg bowel), gastric or endometrial cancer. Use in diagnosis, prognostic assessment, treatment selection, or genetic screening. This application provides for the first time a plasma MSI detection method that can determine the microsatellite (MS) status of a sample with high accuracy and sensitivity.

Description

[0001] field of invention [0002] The present invention relates to biomarker combinations, kits for their detection, and their detection of microsatellite steady state in plasma samples and non-invasive diagnosis of cancer, preferably colorectal cancer (such as bowel cancer), gastric cancer or endometrial cancer, prognosis Evaluation, treatment selection or use in genetic screening. [0003] Background of the invention [0004] Microsatellites are short repetitive DNA sequences or single nucleotide regions contained in genes. In tumor cells, when DNA methylation or gene mutation causes mismatch repair gene loss, it can lead to microsatellite repeat sequence mismatch (microsatellite mutation), resulting in shortening or lengthening of its sequence, thereby causing microsatellite instability ( Microsatellite Instability, MSI). According to the degree of MSI instability, it can be divided into microsatellite instability-high (MSI-H), microsatellite instability-low (MSI-L), and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
Inventor 汉雨生刘成林张之宏张周段飞蝶
Owner GUANGZHOU BURNING ROCK DX CO LTD
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