Generic cancer category multi-gene MSI site targeted detection probe and kit and application

A detection kit and detection probe technology, applied in the field of tumor detection, can solve the problems of few detection sites or proteins, highly dependent detection results, false negative results, etc.

Active Publication Date: 2021-09-10
GUANGZHOU KINGMED TRANSFORMATIVE MEDICINE INST CO LTD +2
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, MSI-PCR and MMR-IHC methods are widely used in clinical practice to detect MSI. However, due to the small number of detection sites or proteins, and the detection results are highly dependent on subjective experience judgments, it is often impossible to accurately predict the MSI status

Method used

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  • Generic cancer category multi-gene MSI site targeted detection probe and kit and application
  • Generic cancer category multi-gene MSI site targeted detection probe and kit and application
  • Generic cancer category multi-gene MSI site targeted detection probe and kit and application

Examples

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Effect test

Embodiment 141

[0042] Example 1 Detection of tumor paraffin-embedded tissues of 41 cancer patients

[0043] The detection probes for the 30 MSI loci with statistically different alleles (p<0.05) between tumors and normal controls determined in this example are as described above.

[0044] 1) Baseline data model establishment

[0045] The MS-NGS method was used to detect the DNA of peripheral blood leukocytes of 30 normal non-cancer populations; the alleles of the MSI loci in the normal non-cancer population (MSS by default) were determined as the baseline reference value. Proceed as follows:

[0046] a. Count the frequency distribution of alleles at each MSI site, where the frequency of a specific allele is expressed as the number of specific allele reads / the number of allele reads with the highest expression frequency, all detected at the MSI site The frequencies of the alleles form a list; for example MSI locus A, with the following allele A 1...A n , and the respective read numbers ar...

Embodiment 2

[0074] Tumor tissues from 6 patients with colorectal cancer were collected, DNA was extracted, and NGS library was constructed. Then use the above probe combination to capture the target MSI site and perform sequencing. Sequencing data is analyzed using this method to determine the status of each MSI site. At the same time, compared with the MSI-PCR method, the results are shown in Table 6, showing that the two methods are consistent.

[0075] Table 6

[0076]

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Abstract

The invention relates to a high-coverage generic cancer category multi-gene MSI site targeted detection probe and kit and application. The generic cancer category multi-gene MSI site targeted detection probe comprises at least two of MSI site detection probes with statistical differences in 30 alleles in a tumor and a normal control. Microsatellites are polymorphic DNA loci containing repeated nucleotide sequences, contain a large number of homopolynucleotide sequences, are extremely unbalanced in base distribution, are high in capture difficulty and have high design requirements on the probes. When the generic cancer category multi-gene MSI site targeted detection probe is designed, the difference of the target MSI in the tumor and normal tissues, the coverage degree in capture sequencing and the capture specificity of a target fragment are fully considered. The length of the probe ranges from 80 bp to 120 bp, the longer the probe is, the higher the fault tolerance is, the higher the MSI capture efficiency is, and meanwhile, higher specificity is achieved. Meanwhile, for MSI sites with high repeatability, a head-to-tail type double-probe design is adopted, so that the capture efficiency can be effectively improved.

Description

technical field [0001] The invention relates to the field of tumor detection, in particular to a pan-cancer multigene MSI site-targeted detection probe, kit and application. Background technique [0002] Microsatellites are polymorphic DNA loci containing repetitive nucleotide sequences, usually 2 to 7 nucleotides per unit. Microsatellite instability (MSI) refers to the phenomenon of changes in the length of microsatellite sequences caused by mutations such as insertions or deletions during DNA replication, and is often related to deficient mismatch repair (dMMR). According to the degree of MSI instability, it can be divided into microsatellite sequence high instability (MSI-High, MSI-H), microsatellite sequence low instability (MSI-Low, MSI-L) and microsatellite stability (microsatellite stability, MSS). Tumors with a high incidence of MSI-H include endometrial cancer, colon adenocarcinoma, gastric adenocarcinoma, and rectal adenocarcinoma. [0003] Study finds that MSI ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11C40B40/06
CPCC12Q1/6886C40B40/06C12Q2600/156C12Q2600/112
Inventor 黄晓强欧小华袁杰铖范喜杰蒙裕欢李桂彬缪夏萍于世辉程雅婷
Owner GUANGZHOU KINGMED TRANSFORMATIVE MEDICINE INST CO LTD
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