Method and system for NGS calibration of lung cancer targeted drug and chemotherapy drug genomes by DNA and RNA double library construction

A DNA library and genome technology, applied in the field of gene sequencing, can solve the problems of low detection efficiency, inability to detect genetic variation, and high detection cost

Pending Publication Date: 2021-01-15
合肥金域医学检验实验室有限公司
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  • Abstract
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Problems solved by technology

Targeted drug therapy through lung cancer drug gene detection can significantly improve the chemotherapy effect, accuracy, and drug safety. The existing lung cancer drug gene detection system focuses on the detection of a single targeted drug or chemotherapy drug gene, and individual targeting genes have also emerged. Mutation detection combination, but it is mainly designed for pan-cancer types, and is limited to the detection of conventional mutations such as point mutations. It cannot detect disease-related gene mutations caused by gene fusion mutations and splicing mutations. The detection efficiency is low and the detection cost is high.

Method used

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  • Method and system for NGS calibration of lung cancer targeted drug and chemotherapy drug genomes by DNA and RNA double library construction
  • Method and system for NGS calibration of lung cancer targeted drug and chemotherapy drug genomes by DNA and RNA double library construction
  • Method and system for NGS calibration of lung cancer targeted drug and chemotherapy drug genomes by DNA and RNA double library construction

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Embodiment 1

[0079] Such as figure 1 The method shown here provides a DNA and RNA dual library construction method for NGS calibration of the genome of lung cancer targeted drugs and chemotherapy drugs: by directly amplifying the genes of known mutation sites and then building a library, and performing NGS sequencing detection, Combined with bioinformatics analysis to determine and verify the existence of the mutant gene; specifically include the following steps:

[0080] S1: Nucleic acid extraction: extract DNA according to "FFPE Tissue Genomic DNA Extraction Standard Operating Procedure", extract RNA according to "Tissue RNA Extraction Standard Operating Procedure", and measure the concentration of DNA / RNA;

[0081] The concentration operating method of described measuring DNA / RNA is: use dsDNA / RNA HS Assay Kit ( dsDNA / RNA High Sensitivity Kit) and 3.0 Fluorometer to measure the DNA / RNA concentration, the specific operation is as follows:

[0082] a) Add 1 μL Qubit Reagent t...

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Abstract

The invention discloses a method and system for NGS calibration of lung cancer targeted drug and chemotherapy drug genomes by DNA and RNA double library construction. The method comprises the following steps: directly amplifying genes with known mutation sites, then constructing a library, performing sequencing detection on NGS, and determining and verifying the existence of mutant genes in combination with bioinformatics analysis. The method comprises the following steps: S1, extracting nucleic acid, and determining the concentration of DNA / RNA; S2, constructing a DNA or RNA library; S3, performing on-machine sequencing: loading a high-throughput sequencing library on a machine, operating equipment, and performing NGS sequencing; S4, performing data bioinformatics processing and analysis:uploading a Fastq format file obtained by sequencing to a bioinformatics analysis software system matched with a true solid organism for automatic analysis; and S5, performing quality control. According to the invention, the problem that the existing lung cancer drug gene detection system can only aim at a targeted drug or a chemotherapeutic drug is solved, lung cancer chemotherapy and targeted drug gene detection are realized in one project, the detection cost is low, and the detection efficiency is high.

Description

technical field [0001] The invention belongs to the technical field of gene sequencing, and in particular relates to a method and a system for DNA and RNA dual library construction for NGS calibration of lung cancer targeting drug and chemotherapy drug genome. Background technique [0002] Tumor is a complex disease caused by the alteration of multiple genes, which leads to the dysfunction of related genes, and the alteration of multiple genes is also the most fundamental reason for the occurrence and development of tumors. Cancer patients have obvious individual differences in various drugs, which are also related to the differential expression and polymorphism of individual tumor-related genes. The mutation status of tumor-related genes plays a very important role in the diagnosis, treatment, prognosis and other personalized treatment of tumors. As more and more markers are identified, the treatment of tumors will be based on patient-specific Biomarkers are becoming more ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C12Q1/6806C12N15/10C40B50/06C12Q1/6886
CPCC12Q1/6869C12Q1/6806C12N15/1096C40B50/06C12Q1/6886C12Q2600/156C12Q2600/158C12Q2600/106C12Q2535/122C12Q2563/107C12Q2531/113C12Q2523/308C12Q2563/143C12Q2563/149
Inventor 李志强李晓华陈浩
Owner 合肥金域医学检验实验室有限公司
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