Tumor-related microsatellite instability site combination and application thereof

A technology of microsatellite instability and microsatellite loci, applied in the field of microsatellite instability locus combination, can solve the problems of inconsistent antibodies and inconsistent reading standards of pathologists, so as to reduce the detection cost and reduce the combination of MS loci. , the effect of low cost

Inactive Publication Date: 2022-01-28
BEIJING USCI MEDICAL DEVICES CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using IHC to assess MMR protein levels has the problems of inconsistent use of antibodies and inconsistent reading standards for pathologists

Method used

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  • Tumor-related microsatellite instability site combination and application thereof
  • Tumor-related microsatellite instability site combination and application thereof
  • Tumor-related microsatellite instability site combination and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Obtaining combination of tumor-associated microsatellite instability sites

[0028] Include the following steps:

[0029] 1. Scan 2,952,815 MS sites with a repeat unit of 1-5 bases and a repeat length of 10-100 bases available on the human reference genome.

[0030] 2. Since the use of single-base repeat MS sites has higher sensitivity than the use of poly-base repeat MS sites, 2263 single-base repeat MS sites in step 1 were screened out.

[0031] 3. Select 363 MS sites covered by the panel for detecting cancer driver mutations and drug target mutations.

[0032] 4. In the training set samples of 28 cases of primary colorectal cancer (7 cases of microsatellite instability (MSI-H), 21 cases of microsatellite stable (MSS)), use MANTIS software to calculate 363 MS bits in step 3 The MSI of the points is scored MSI-score; the calculation method of MSI-score is to determine the stability of 363 sites in each sample, and calculate the proportion of unstable sites ...

Embodiment 2

[0038] Example 2 The application of the site combination of tumor-related microsatellite instability

[0039] Using the MSI-PCR results as the gold standard, the performance of using the 9 sites in Table 1 to determine MS status was evaluated.

[0040] 1. DNA was extracted from tumor tissues and paracancerous tissues of 36 patients with primary colorectal cancer (7 cases of MSI, 29 cases of MSS):

[0041] 1) Take a mung bean-sized fresh tissue, cut it into pieces with a disposable blade and add it to a centrifuge tube;

[0042] 2) Add proteinase K and lysate to the tube respectively, incubate overnight at 56°C until the sample is completely lysed;

[0043] 3) Add buffer solution and absolute ethanol and mix well, transfer to the adsorption column, add rinse solution, centrifuge, discard the waste solution, and elute the salt;

[0044] 4) Transfer the adsorption column to a new collection tube, add eluent, and collect the DNA.

[0045] 2. Use a 2.2M panel containing 9 MS sites...

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Abstract

The invention provides a set of tumor-related microsatellite instability site combination and application thereof. The tumor-related microsatellite instability site combination comprises nine microsatellite loci (SEQ ID NO: 1-9) from MS1 to MS9. The nine microsatellite loci provided by the invention are based on an NGS technology, so that the defects of subjective manual misjudgment, complicated operation, high cost and the like existing in methods such as IHC and MSI-PCR can be effectively avoided. The nine MSI loci are additionally added in an original detection Panel, MSI can be detected only through one-time sampling, and information such as tumor mutation load, embryonic system mutation and system mutation can be obtained. By using the 9 loci, MS detection can be carried out with relatively high sensitivity and specificity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the site combination and application of tumor-related microsatellite instability. Background technique [0002] Microsatellite (MS), also known as short tandem repeat sequence, is a short repetitive nucleotide sequence with a unit length between 1 and 6 bases, which widely exists in eukaryotic genomes. Microsatellite instability (MSI) indicates the insertion or deletion of nucleotides in a microsatellite locus. Changes in microsatellite regions typically occur during DNA replication. DNA mismatch repair (MMR) proteins, such as MLH1, MSH2, MSH6, and PMS2, are responsible for the repair of MSI. Germline or somatic inactivation of MMR genes may in turn lead to MSI. MSI is found in many cancer types, with the highest incidence in colon and endometrial cancers. Germline mutations in MMR proteins are associated with the pathogenesis of Lynch syndrome, which accounts for approximately 2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/156
Inventor 姬晓勇徐冰单光宇伍启熹王建伟
Owner BEIJING USCI MEDICAL DEVICES CO LTD
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