Next generation sequencing-based detection panel and detection kit for pan-cancer targeting, chemotherapy and immune drugs and application thereof

A detection kit and next-generation sequencing technology, applied in the field of biomedicine

Active Publication Date: 2019-04-12
ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The present invention aims to provide a detection panel, a detection kit and its application for pan-cancer targeting, chemotherapy and immunotherapy based on next-generation sequencing, so as to solve the problem of not being able to comprehensively detect samples in the prior art. technical problem

Method used

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  • Next generation sequencing-based detection panel and detection kit for pan-cancer targeting, chemotherapy and immune drugs and application thereof
  • Next generation sequencing-based detection panel and detection kit for pan-cancer targeting, chemotherapy and immune drugs and application thereof
  • Next generation sequencing-based detection panel and detection kit for pan-cancer targeting, chemotherapy and immune drugs and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The panel of this example includes gene mutations related to pan-cancer type, treatment, and prognosis, microsatellite instability sites, and exon regions related to tumor mutation load calculation. Correspondingly, the kit of this embodiment includes detection probes, and the detection probes target gene mutations related to pan-cancer type, treatment, and prognosis, and exon regions related to microsatellite instability sites and tumor mutation load calculation. , detection probe coverage area / target area >= 99%. Detection probes are designed according to conventional design rules in the art.

[0043]Among them, gene mutations related to pan-cancer type, treatment, prognosis and exon regions related to tumor mutation burden calculation include: ABCA13, ABCA8, ABL1, ACADSB, ACOT13, ADAMTS6, ADRB1, ADSS, AGPAT9, AK7, AKT1, AKT2, AKT3, ALG9, ALK, ALK_fus, ALOX12B, ALS2CR11, AMER1, ANKRA2, ANKRD46, ANO1, APC, APOPT1, AR, ARAF, ARHGAP4, ARHGAP6, ARID1A, ARID1B, ARID2, ARI...

Embodiment 2

[0046] Use the kit in Example 1 to detect the standard substance of simulated tissue DNA.

[0047] 1. Preparation of verification standards

[0048] Twenty mutant standard DNAs were prepared by mixing proportionately the DNA of the tumor cell line containing the known mutation and the DNA of the wild-type cell line GM12878. And the frequency of each mutation in the standard was determined by ddPCR. Tumor cell line DNA is shown in Table 1.

[0049] Table 1

[0050]

[0051]

[0052] 2. DNA interruption:

[0053] DNA was quantified using Qubit 3.0 and the dsDNA HS Assay Kit.

[0054] Cut the polytetrafluoroethylene wire to a length of about 1 cm with UV-sterilized medical scissors, and ensure that the length of the interrupted rod is well uniform, place it in a clean container, and sterilize it with UV light for 3 to 4 hours. After the sterilization is completed, a 1 cm polytetrafluoroethylene thread is loaded into a 96-well plate with sterilized tweezers. Put 2 inte...

Embodiment 3

[0148] The library construction and capture of cfDNA samples were performed using the kit of Example 1 of the present invention.

[0149] 1. Preparation of verification standards

[0150] Twenty mutant standard DNAs were prepared by mixing proportionately the DNA of the tumor cell line containing the known mutation and the DNA of the wild-type cell line GM12878. And the frequency of each mutation in the standard was determined by ddPCR. Tumor cell line DNA is shown in Table 1. The standard DNA was interrupted to mimic the fragment length of cfDNA (~165 bp).

[0151] 2. Library Construction

[0152] 1. End repair and add A tail at the 3' end:

[0153] 1.1 Take 50 μL of cfDNA, make up to 50 μL with nuclease-free water if it is less than 50 μL, and add it to the reaction system according to Table 2 in Example 2.

[0154] 1.2 Mix by vortexing, microcentrifuge, and place in a PCR machine. The reaction procedure is as shown in Table 3 in Example 2.

[0155] 2. Connect the conn...

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Abstract

The invention discloses a next generation sequencing-based detection panel and detection kit for pan-cancer targeting, chemotherapy and immune drugs and application thereof. The detection panel includes gene mutations related to pan-cancer type, treatment and prognosis, tumor mutation load calculation related exon regions and microsatellite instability sites. According the technical solution of the present invention, the detection panel includes the gene mutations related to pan-cancer type, treatment and prognosis, the tumor mutation load calculation related exon regions and the microsatellite instability sites, and the gene information included is comprehensive; a variety of tumor mutations can be jointly detected; and the detection panel and detection kit can be used for the concomitantdiagnosis of targeted drugs, chemotherapeutics or immune drugs to obtain accurate results.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular, to a detection panel, a detection kit and application thereof for pan-cancer species targeting, chemotherapy and immunotherapy based on second-generation sequencing. Background technique [0002] Currently, there are many cancer-related detection technologies, such as high-throughput sequencing, target region capture sequencing, liquid biopsy, tumor mutational burden, and MSI detection. [0003] Among them, High-Throughput Sequencing (High-Throughput Sequencing), also known as Next Generation Sequencing (NGS), is relative to traditional Sanger Sequencing (Sanger Sequencing). [0004] Based on the sequencing method of Sanger et al., through technological innovation, the fragmented genomic DNA is connected with adapters on both sides, and then different methods are used to generate millions of space-fixed PCR clone arrays. Each clone consists of multiple copies of a sing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/106C12Q2600/118C12Q2600/112C12Q2600/156
Inventor 洪媛媛夏艳颜林林闫慧婷宋小凤曾雪霞郭现超赵利利朱伟何骥杜波陈维之
Owner ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD
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