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Method for directly detecting P53 gene mutation in lung cancer sample based on nano probe

A nano-probe and sample technology, applied in the direction of biochemical equipment and methods, microbial measurement/testing, etc., to achieve the effects of low background, easy analysis, and improved hybridization specificity

Active Publication Date: 2009-03-25
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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Problems solved by technology

[0009] The purpose of the present invention is to provide a method for directly detecting P53 gene mutation in lung cancer samples based on nanoprobes. The technical problem to be solved by the present invention is to provide a Detection method of nanoprobe chip for detecting P53 gene mutation in lung cancer patients based on "sandwich hybridization" principle

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  • Method for directly detecting P53 gene mutation in lung cancer sample based on nano probe
  • Method for directly detecting P53 gene mutation in lung cancer sample based on nano probe
  • Method for directly detecting P53 gene mutation in lung cancer sample based on nano probe

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Embodiment Construction

[0041] The present invention will be further described below through embodiment and accompanying drawing.

[0042] 1. Prepare materials and reagents

[0043] (1) EasyHtyb hybridization solution: purchased from Roche Company;

[0044] (2) All probes were synthesized in Dalian Bao Biological Engineering (Dalian) Co., Ltd. (Dalian TaKaRa Company)

[0045] (3) 15nm and 30nm gold nanoparticles were purchased from Shanghai Water Source Biotechnology Co., Ltd.

[0046] (4)NaNO 3 Lotion: 1M NaNO3 300ml

[0047] 0.2M Na 2 HPO 4 30.5ml

[0048] 0.2M NaH2PO4 9.5ml

[0049] Make up to 1000ml

[0050] (5) PB solution: 0.3mol / L NaNO3 and 10mmol / L Na2HPO4 / NaH2PO4, pH 7.0

[0051] (6) Silver stain enhancement solution: 60 μl of 1% gelatin

[0052] 2.55g citric acid / 2.35g; trisodium citrate / 10ml H 2 O; 10 μl

[0053] 1.7g hydroquinone / 30ml H 2 O, 30 μl

[0054] 0.5g AgNO 3 2ml H 2 O; 1....

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Abstract

The invention relates to a method for directly detecting p53 gene mutation in lung cancer samples based on nanoprobe. The method is characterized in that sandwich hybridization is carried out to an unmarked target nucleic acid sequence, a signal probe marked by nano gold and a capture probe connected with a solid-phase holder and the amplification effect of silver staining enhanced nano-gold signals is used for generating highly sensitive identification signals. The method can detect single base mutation in non-amplified genomic DNA samples. Compared with traditional method based on fluorescence signal detecting, the nano chip detecting method greatly enhances the sensitivity and specificity of detection and hybridization signals can be collected and analyzed by an ordinary optical scanner or a charge-coupled device (CCD) digital camera or analyzed by related software to obtain detecting reports.

Description

technical field [0001] The invention relates to a method for directly detecting P53 gene mutation in lung cancer samples based on a nano-probe, more precisely, the invention relates to a nano-diagnosis chip for detection of P53 gene mutation for auxiliary diagnosis of lung cancer. In particular, a nano-probe chip detection method for directly detecting gene mutations in genomic DNA. Background technique [0002] Lung cancer is one of the malignant tumors with high morbidity and mortality in the world, and its case fatality rate ranks first among the malignant tumor deaths of the urban population. Although the treatment of lung cancer has made great progress in the past 20 years, the prognosis of lung cancer patients has not improved significantly, and the 5-year survival rate is only 10% to 15%. The main reason for this is that routine screening programs for lung cancer are inadequate and fail to identify early lesions of cancerous development. [0003] With the developmen...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 毛红菊贾春平金庆辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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