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System capable of simultaneously detecting tumor targeting therapy related target spots and immunotherapy related TMB and MSI

An immunotherapy and tumor targeting technology, applied in biochemical equipment and methods, microbial assay/test, genomics, etc., it can solve the problem that local tissue cannot represent the overall situation, strong heterogeneity of tumor tissue, and inability to obtain tumor tissue To improve the positive detection rate, accurate and reliable results, and increase the probability of patient benefit

Pending Publication Date: 2019-07-09
AMOY DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In clinical practice, most tumor surgical tissues or biopsy specimens are used as the detection target. Generally, pathologists can make the tissues into paraffin-embedded slides, and conduct pathological diagnosis on the tumor cell content and tumor type through the morphology of tissue cells. The disadvantage lies in the existence of tumor tissue. Strong heterogeneity, local tissue cannot represent the overall situation, and many patients with advanced cancer cannot obtain tumor tissue, losing many treatment options
[0007] To sum up, there are currently many molecular targets clinically used for tumor treatment, involving multiple genes and multiple mutation types within the genome. Traditional detection methods are diverse, but they can only detect individual targets. Accurate and comprehensive Treatment requires the comprehensive use of multiple methods and multiple technology platforms, with high cost, slow speed, and complicated operation

Method used

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  • System capable of simultaneously detecting tumor targeting therapy related target spots and immunotherapy related TMB and MSI
  • System capable of simultaneously detecting tumor targeting therapy related target spots and immunotherapy related TMB and MSI
  • System capable of simultaneously detecting tumor targeting therapy related target spots and immunotherapy related TMB and MSI

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Detection of known hotspot mutation sample sites

[0039] Detection steps:

[0040] 1. DNA fragmentation: Take 200ng of DNA from FFPE samples that pass the test, and use a 130ul volume fragmentation tube for fragmentation. After fragmentation, take 125ul fragmentation products, use 100ul magnetic beads and 50ul magnetic beads for fragment screening, and 25ul purified water Perform elution.

[0041] 2. Library preparation: Quantify the fragmented DNA, subject to the fluorescence quantitative method, take 20ng of the fragmented product, add water to 25ul, add 3.5ul end repair buffer and 1.5ul end repair enzyme, run according to the following procedure: 20 Store at ℃30min+65℃30min+4℃. Add 0.3ul linker, 15ul ligation buffer and 0.5ul enhancer to the end-repair product, store at 20°C for 15min, and then store at 4°C. The ligation product was purified with 44ul magnetic beads and eluted with 25ul purified water. Take 23ul of ligated purified product, add 25ul o...

Embodiment 2

[0059] Example 2: 37 samples were tested for TMB, and the operation steps were the same as in Example 1. The results of TMB analysis were compared with the results of WES, and the consistency was high. The data quality control results are shown in Appendix 1, Table 4, and the TMB numerical results are shown in Appendix 1, Table 5.

Embodiment 3

[0060] Example 3: 20 samples were tested by MSI, and the operation steps were the same as those in Example 1. The MSI test results were compared with the Sanger test results, and the consistency was high. The data quality control results are shown in Table 6 of Appendix I.

[0061]

[0062]

[0063] Appendix 1

[0064] Table 1 List of genes used to detect point mutations and insertion-deletion mutations

[0065]

[0066]

[0067] Table 2 Gene list for simultaneous detection of point mutations, indels and fusion mutations

[0068]

[0069] Table 3 Sequencing Data Quality Control Information Table 1

[0070]

[0071]

[0072] Table 4 Sequencing Data Quality Control Information Table 2

[0073]

[0074]

[0075] Table 5 TMB value information table

[0076]

[0077]

[0078] Table 6 Sequencing Data Quality Control Information Table 3

[0079]

[0080] Appendix II

[0081] Table 2 Microsatellite locus information

[0082]

[0083]

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Abstract

The invention discloses a system capable of simultaneously detecting tumor targeting therapy related target spots and immunotherapy related TMB and MSI. The system comprises a library preparation reagent, a probe, a hybridization capture reagent and a data analysis system, wherein the library preparation reagent, the probe and the hybridization capture reagent can be used for sequencing a library,and the data analysis system can analyze results including data quality control information and statistical information, mutation site information and TMB values.

Description

[0001] Technical field: [0002] The invention relates to a gene detection system developed based on a high-throughput sequencing platform, which can simultaneously detect gene information related to tumor-related targeted therapy and immunotherapy. [0003] Background technique: [0004] A large number of clinical studies have shown that targeted therapy drugs represented by TKI small-molecule inhibitors have brought more benefits to cancer patients, and corresponding companion diagnostic products have become a must-choice item before formulating clinical treatment plans. one. Although targeted therapy can benefit patients more, more than 30% of patients cannot choose this treatment because they do not meet the criteria for targeted therapy. In recent years, breakthroughs in immune checkpoint inhibitor drugs have brought new hope to cancer patients, especially the clinical research of PD1 / PDL1 drugs is the most popular, but it is not recommended to be used in patients with dr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6806G16B30/00G16B20/50
CPCC12Q1/6886C12Q1/6806C12Q2600/156C12Q2600/106C12Q2525/191
Inventor 葛会娟田晓岩李旭超刘炜芬刘志源黄可君李健鹏杨爽吴建明
Owner AMOY DIAGNOSTICS CO LTD
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