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Primers and probe for detecting EGFR gene mutation, kit thereof, and application method of kit

A technology for detection kits and probes, applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., can solve problems such as quantitative discrimination of gene mutations

Active Publication Date: 2013-01-02
上海源奇生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Patent ZL200910111499.2 discloses a primer and probe for detecting human EGFR gene mutations, and also discloses a kit containing the primers and probes. The kit can only detect the following 29 EGFR mutations and cannot detect The type of mutation, especially the mutation type of exon 19 of the EGFR gene, is analyzed
In addition, it is impossible to judge the amount of gene mutation

Method used

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  • Primers and probe for detecting EGFR gene mutation, kit thereof, and application method of kit
  • Primers and probe for detecting EGFR gene mutation, kit thereof, and application method of kit
  • Primers and probe for detecting EGFR gene mutation, kit thereof, and application method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0146] Kit composition:

[0147] Table 9:

[0148]

[0149]

Embodiment 2

[0150] Embodiment 2 adopts the kit of embodiment 1 to detect:

[0151] A. PCR fluorescence detection:

[0152] 1. Sample processing:

[0153] Samples 1-12 were taken, extracted with QIAGEN Paraffin Tissue DNA Extraction Kit or QIAGEN Whole Blood DNA Extraction Kit, and stored at -20°C.

[0154] Specimen 1-3: Paraffin specimen;

[0155] Specimen 4-6: fresh specimen;

[0156] Specimen 7-9: Peripheral blood specimen;

[0157] Specimen 10-12: peripheral plasma specimen;

[0158] 2. Amplification reagent preparation:

[0159] The corresponding PCR reaction solution was taken out from the kit, melted at room temperature, shaken and mixed, and then centrifuged at 2000rpm for 10s.

[0160] Pack 21ul of reaction solution and 1ul of enzyme solution in each tube into PCR reaction tubes; note that the number of PCR tubes for each reaction solution should be the sum of the number of samples, one negative control and one positive control tube;

[0161] 3. Adding samples: Take 3ul fro...

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Abstract

The invention relates to the field of gene mutation detection, and specifically relates to primers and a probe used for detecting EGFR gene mutation, a kit thereof, and an application method of the kit. With the kit provided by the invention, a sample containing 0.1-1% of EGFR gene mutation DNA can be detected; and accurate genotyping can be carried out upon a sample containing 10% of EGFR No. 19 exon gene mutation DNA through sequencing. The sensitivity of the kit provided by the invention is high, wherein a minimum detection limit is 5-copy gene. With the kit, qualitative detection can be carried out upon EGFR gene mutation. Different sensitivity reference ranges can be provided for assisting clinical determinations of mutation amount volumes, such that final treatment programs can be provided according to different mutations.

Description

technical field [0001] The invention relates to the field of gene mutation detection, in particular to a primer, a probe, a kit and a use method for detecting EGFR gene mutation. Background technique [0002] Traditional chemotherapy mainly targets the DNA of cells. This effect often lacks specificity. While killing tumor cells, it also "wrongly kills" many normal cells. Targeted therapy is completely different from previous chemotherapy. There are many mechanisms involved in the pathogenesis of tumors, and these mechanisms are often unique to tumor cells. Targeted therapy is aimed at a specific signal transduction pathway or a certain pathogenesis in the pathogenesis of tumors, using monoclonal antibodies and small molecular substances to interrupt it, so as to achieve the purpose of treating tumors, but has a slight effect on normal cells. [0003] Targeted therapy is highly specific and only targets cancer cells. It does not have the common side effects of chemotherapy,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 熊慧程新建谢立群陈嘉铮包文静陶慧卿丁璐徐任占闽宁杨青喜
Owner 上海源奇生物医药科技有限公司
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