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Screening method and screening device of microsatellite sites pertinent to genome stability, and application

A genome stability, microsatellite locus technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc., can solve the problem of low accuracy of test results, achieve high accuracy, sensitivity and highly specific effect

Active Publication Date: 2020-04-03
BEIJING USCI MEDICAL LAB CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The main purpose of the present invention is to provide a screening method, screening device and application of microsatellite loci related to genome stability, to solve the problem of detection in the prior art The problem of low accuracy of results

Method used

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  • Screening method and screening device of microsatellite sites pertinent to genome stability, and application
  • Screening method and screening device of microsatellite sites pertinent to genome stability, and application
  • Screening method and screening device of microsatellite sites pertinent to genome stability, and application

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Embodiment 1

[0047] In a preferred embodiment of the present application, a method for screening microsatellite sites related to genome stability is provided, figure 1 is a flowchart of a method according to an embodiment of the present invention. As shown, the screening method includes:

[0048] Step S101, using the first method to measure the state of microsatellite sites and sample state of tumor tissues and blood cells in multiple samples;

[0049] Step S102, using a second method different from the first method to verify the sample states of multiple samples;

[0050] Step S103, selecting a plurality of samples with the same sample status determined by the first method and the second method, and according to the sample status of each sample, classifying the multiple samples with the same sample status into the first type of MSS (i.e. MSS) samples and MSI-H (MSI-H) of the second category of samples;

[0051] Step S104, respectively selecting a plurality of microsatellite loci with ...

Embodiment 2

[0066]This embodiment provides a method for screening colorectal cancer tissue-specific microsatellite loci, and using this method to detect and verify the status of the samples. Specific steps are as follows:

[0067] 1. Obtain tumor tissue and white blood cell DNA from several colorectal cancer patients.

[0068] 2. Use the high-throughput microarray capture method to obtain the genome sequence of patient sample tumor and leukocyte control sample.

[0069] 3. Align the sequence obtained by sequencing with the human hg19 reference genome sequence, and remove the repeated sequence and the sequence aligned to multiple positions in the alignment result.

[0070] 4. Use the MSIsensor tool to scan the human hg19 reference genome sequence to obtain the distribution of microsatellite loci within the human genome.

[0071] 5. From the microsatellite sites distributed in the whole human genome, select all the microsatellite sites covering the capture interval of the chip.

[0072] ...

Embodiment 3

[0084] In this embodiment, a microsatellite site for detecting genome stability is provided, and the microsatellite site is screened by any of the above screening methods. The microsatellite sites screened by the above method of the present application are highly correlated with the unstable state of the microsatellite, so the stability state of the sample to be tested can be accurately distinguished, and the detection sensitivity and specificity are high (verified by MSI-PCR 64 cases of colorectal cancer samples, including 14 cases of MSI-H samples, 50 cases of MSS samples, using the microsatellite loci screened by the method of this application for genome stability detection, the method is correctly detected, sensitivity and specificity All of them are 100%, see Table 2 below for details.

[0085] Table 2:

[0086]

[0087] In a preferred embodiment, the microsatellite locus is a marked microsatellite locus of colorectal cancer tissue, and preferably the microsatellite l...

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Abstract

The invention provides a screening method and screening device of microsatellite sites pertinent to genome stability, and an application. The screening method comprises the steps of determining the state of microsatellite sites of respective tumor tissue and blood cells of a plurality of samples and the sample state of the samples by a first method; performing verification on the sample state of the samples by a second method; selecting the samples consistent in the sample state, determined by the first method and the second method, and dividing the samples consistent in the sample state, intoMSS first-class samples and MSI-H second-class samples; respectively screening out a first site set formed by a plurality of microsatellite sites highest in relevance with the first-class samples, and a second site set formed by a plurality of microsatellite sites highest in relevance with the second-class samples; and taking the intersection of the first site set and the second site set to obtain the microsatellite sites pertinent to the genome stability. The microsatellite sites screened by the method are high in detection accuracy and high in detection sensitivity and specificity.

Description

technical field [0001] The invention relates to the field of gene sequencing data analysis, in particular to a screening method, screening device and application of microsatellite sites related to genome stability. Background technique [0002] Microsatellite loci are generally short repetitive sequences with a length of about 1-6 bp, which widely exist in the human genome. Microsatellite instability refers to any change in the length of a microsatellite in a tumor due to insertion or deletion of repeating units compared with normal tissue, and new microsatellite alleles appear. The mechanism of its occurrence mainly includes the slippage of DNA polymerase leading to the mismatch of one or more bases in the repeat sequence and the deletion or insertion of base pairs caused by microsatellite recombination. [0003] The traditional detection method of microsatellite instability mainly uses MSI-PCR to detect the stability of five microsatellite loci (NR-27, NR-24, NR-21, BAT-2...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/686C12Q2600/156C12Q2525/151
Inventor 张静波单光宇李孟键伍启熹王建伟刘倩唐宇
Owner BEIJING USCI MEDICAL LAB CO LTD
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