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Genetic mark, primer, probe and kit for detecting lung cancer

A technology for detecting kits and gene markers, which is used in the field of probes, kits, primers, and gene markers for detecting lung cancer, which can solve the need for the diagnosis of lung cancer markers, and is not suitable for early detection or multiple inspections of lung cancer, etc. problems, to avoid excessive tissue biopsy, improve reproducibility and accuracy, reduce medical costs and expenses

Active Publication Date: 2016-06-15
王义明
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The detection of lung cancer markers in tissue requires surgery or biopsy to obtain cancer tissue, which is not suitable for early detection or multiple inspections of lung cancer, because once biopsy or surgery is required, lung cancer is already in a relatively advanced stage, and the diagnosis of lung cancer markers is no longer necessary

Method used

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  • Genetic mark, primer, probe and kit for detecting lung cancer
  • Genetic mark, primer, probe and kit for detecting lung cancer
  • Genetic mark, primer, probe and kit for detecting lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Preparation and use of the kit for detecting lung cancer

[0044] 1. The composition of the real-time fluorescent quantitative PCR kit for detecting the expression levels of SFTPB, FGFR1 and FILIP1L genes:

[0045] SFTPB gene specific primer (upstream and downstream primer mix), 10 μM; specific fluorescent probe, 2 μM;

[0046] FGFR1 gene-specific primers (upstream and downstream primer premix), 10 μM; specific fluorescent probes, 2 μM;

[0047] FILIP1L gene-specific primers (upstream and downstream primer mix), 10 μM; specific fluorescent probes, 2 μM;

[0048] GAPDH gene-specific primers (premixed upstream and downstream primers), 10 μM; specific fluorescent probes, 2 μM;

[0049] UniversalPCRMasterMix, purchased from ABI company;

[0050] RNase-freewater was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.

[0051] Positive control substances were stored separately, and were diluted in concentration gradients when used.

[0052] 2. H...

Embodiment 2

[0066] Embodiment 2 clinical sample detection

[0067] 1. Clinical sample collection:

[0068] A total of 50 samples from healthy volunteers and 50 samples from lung cancer patients were collected in this study.

[0069] 2. Extraction of total RNA from fresh peripheral blood:

[0070] (1) 1 mL of whole blood anticoagulated with EDTA (ethylenediaminetetraacetic acid), centrifuge at 5000 rpm for 10 min, discard the plasma, add 1 mL of erythrocyte lysate, mix well, put it on ice for 15 min, and centrifuge at 5000 rpm for 5 min;

[0071] (2) Discard the supernatant, add 1 mL of PBS (Phosphate Buffered Saline, phosphate buffered saline), mix well, and centrifuge at 5000 rpm for 5 min; repeat the operation three times until the supernatant is transparent, discard the supernatant;

[0072] (3) Add 1mL Trizol reagent to the sedimented cells in the lower layer, mix well, and place at room temperature for 5 minutes;

[0073] (4) Add 200 μL of chloroform, mix thoroughly for 30 seconds...

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Abstract

The invention discloses a genetic mark for detecting lung cancer. The genetic marker comprises an SFTPB gene, an FGFR1 gene and an FILIP1L gene. Research studies show that mRNAs (messenger ribonucleic acids) of SFTPB, FGFR1 and FILIP1L in blood mononuclear cells of patients with the lung cancer are not expressed normally, accuracy rate of diagnosis on the lung cancer reaches 98.3% after integration, and the genetic mark is much more superior to a single gene in terms of diagnosis specificity and sensitivity. Through measurement of expression quantities of the SFTPB gene, the FGFR1 gene and the FILIP1L gene in blood, diagnosis on the lung cancer (especially early lung cancer from which the patients suffer without symptoms) is achieved, so that clinical lung cancer detection rate is increased, and death rate of the patients with the lung cancer is reduced.

Description

technical field [0001] The invention relates to the technical field of lung cancer detection, in particular to gene markers, primers, probes and kits for detecting lung cancer. Background technique [0002] Lung cancer occurs in the epithelium of the bronchial mucosa. In recent years, the incidence and mortality of lung cancer have been on the rise, and it has become a serious health problem facing the world in the 21st century. In China, it is estimated that by 2025, the number of lung cancer patients will reach 1 million, ranking first in the world. Research on the occurrence, development, treatment and prognosis of lung cancer has always been an important topic facing the medical community. At present, the early diagnosis of lung cancer includes chest X-ray and chest CT examination, sputum cell examination, fiberoptic bronchoscopy and percutaneous lung biopsy. Due to the high cost of these clinical diagnostic methods for lung cancer diagnosis and the specificity of instr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6886C12Q2600/158C12Q2531/113C12Q2561/101C12Q2545/101
Inventor 王义明罗国安
Owner 王义明
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