Kit and use thereof

A kit and probe technology, applied in the field of biomedicine, can solve problems such as low sensitivity, large limitations, and inability to detect unknown mutations, and achieve the effects of high quality, elimination of errors, and improved accuracy

Inactive Publication Date: 2017-07-04
BGI GUANGZHOU MEDICAL LAB CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The PCR-Sanger method has good accuracy and can detect new point mutation types and InDel, but the sensitivity is very low, and it can only detect high-frequency mutations with a mutation rate greater than 20%, and it is difficult to detect multi-gene and multi-site detection
The PCR-Mass method cannot detect unknown mutations, nor can it detect InDel, and the sensitivity is about 5%. This technology has relatively large limitations
The ARMS-PCR method has good specificity and high sensitivity, and can detect 1%-5% low-frequency mutations, but cannot detect unknown mutations, and it is also difficult to simultaneously detect multiple genes and multiple loci

Method used

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  • Kit and use thereof
  • Kit and use thereof
  • Kit and use thereof

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Embodiment 1

[0057] Obtaining the kit of one aspect of the present invention, and realizing the method and / or device of one aspect of the present invention generally include the design of target region capture probes / chips, micro-sample library building, hybridization on-machine sequencing, and biological information analysis of off-machine data and interpretation of variable data.

[0058] (1) Determination, sequence design and production of the genes selected for the chip.

[0059] First, the top 20 genes with the mutation rate in the gastric cancer classification in the COSMIC database were investigated, and the genes related to gastric cancer medication, important oncogenes and tumor suppressor genes found in gastric cancer were screened out, and the middle and high frequency mutations of gastric cancer were added. Genes to obtain the final gene list of the chip, a total of 17, as shown in Table 1.

[0060] The target area of ​​the chip is calculated by a program independently written...

Embodiment 2

[0096] 1. Sample selection and DNA extraction.

[0097] In the embodiment of the present invention, a formalin-fixed paraffin-embedded (FFPE) gastric cancer tissue sample with a known mutation TP53 gene I195F mutation was selected, and the specific sample information is shown in Table 3.

[0098] table 3

[0099] sample number organization type source mutation site access to SC1 FFPE Patients with poorly differentiated gastric adenocarcinoma TP53-I195F clinical sample

[0100]The I195F mutation of the TP53 gene carried by the SC1 sample was verified by Sanger sequencing. Figure 4 .

[0101] Genomic DNA was extracted using DneasyBlood & Tissue Kit Genome Extraction Kit (Qiagen, catalog number: 69504). The concentration of the extracted DNA was detected by a Qubit 2.0 fluorescence spectrophotometer (Life technologies, product number: Q32866).

[0102] 2. Genomic DNA Fragmentation

[0103] The sample fragmentation uses the Covaris system o...

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Abstract

The invention discloses a kit. The kit comprises a probe, the probe is fixed on a solid matrix or is free in a solution, and the probe can specifically identify at least one part of regions in every gene of seventeen genes in table 1. The invention also provides a use of the kit, a method for constructing a target region sequencing library, a sequencing method, a method for detecting the variation of the target region, and a device for detecting the variation of the target region. The kit and/or the method or the device are/is used to once, simply, conveniently and highly-specifically obtain relevant gene sequences of stomach cancer and accurately detect and analyze the relevant gene sequences in order to make the detection and analysis result used for medication guidance and/or risk predication of the stomach cancer in an auxiliary manner.

Description

technical field [0001] The present invention relates to the field of biomedicine, specifically, to a kit and its use, more specifically, the present invention relates to a kit, the use of the kit, a method for constructing a sequencing library of a target region, a sequencing method, a A method and device for detecting variation in a target region. Background technique [0002] Gastric cancer is one of the most common malignant tumors in the world, ranking second in the global cause of cancer death. In 2012, the number of new patients with gastric cancer in the world was about 950,000, and the number of deaths from gastric cancer was about 700,000. The incidence of gastric cancer is the highest in East Asia. . my country is a country with a high incidence of gastric cancer. According to the data of the National Cancer Center, the incidence and mortality of gastric cancer ranked third among malignant tumors in my country in 2011 [Chen Wanqing, Zheng Rongshou, Zeng Hongmei et...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C40B50/06C40B40/06
CPCC12Q1/6874C12Q1/6886C12Q2600/156C40B40/06C40B50/06C12Q2525/191C12Q2535/122
Inventor 叶明芝费凌娜刘继龙
Owner BGI GUANGZHOU MEDICAL LAB CO LTD
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