A dsDNA-agncs fluorescent probe for HER2 detection and its construction method and application

A fluorescent probe and a construction method technology are applied to a dsDNA-AgNCs fluorescent probe for HER2 detection and its construction and application fields, which can solve the problems of limited wide application, high price and the like, and achieve improved sensitivity, simple operation, Small size effect

Active Publication Date: 2020-12-18
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, electrochemical methods have historically faced stability and reproducibility issues and are expensive
This limits the widespread application of these methods

Method used

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  • A dsDNA-agncs fluorescent probe for HER2 detection and its construction method and application
  • A dsDNA-agncs fluorescent probe for HER2 detection and its construction method and application
  • A dsDNA-agncs fluorescent probe for HER2 detection and its construction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The preparation method of silver nanocluster, the steps are as follows:

[0038] Take 40 μL of DNA1 (50 μM) and 40 μL of DNA2 (50 μM), and add 240 μL of phosphate buffer (20 mM, pH 7.0) to mix well, detwirl at high temperature and then cool to room temperature for hybridization and complementation. Add 12 μL AgNO 3 Shake and mix the aqueous solution (1mM) at room temperature for 15min, then add 12μL of freshly prepared NaBH 4 (1mM) mixed quickly and transferred to a constant temperature water bath at 25°C to react for 2 hours, then its fluorescence can be detected. The sequence number of DNA1 is: 5'-GG GGT GTG GCG ACG-3'. The sequence number of DNA2 is: 5'-ATATT ACCC CAA CCCC CGT CGC CAC ACCCC GGGG AAG GGGT AAT AT-3'.

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Abstract

The invention discloses a dsDNA-AgNCs fluorescent probe for detection of HER2, as well as a construction method and an application thereof. DNA1 and DNA2 are hybridized to obtain the dsDNA, which thenis subjected to an in-situ reduction reaction with silver salt and a reducing agent to form AgNCs probe; wherein the DNA1 is a HER2 aptamer DNA fragment, and the DNA2 contains a DNA fragment being complementary with the DNA1, a template DNA fragment for synthesizing AgNCs, and a DNA fragment being rich in G basic group; the two ends of the DNA2 are complementary DNA fragments. The fluorescent probe has advantages of strong signal, high specificity and sensitivity and the like during detection on the HER2. The synthesis method is simple in steps, low in cost and gentle in conditions, and is beneficial to be promoted in production and application.

Description

technical field [0001] The present invention relates to a fluorescent probe for the detection of the biomarker HER2, in particular to a dsDNA-AgNCs fluorescent probe synthesized by hybridizing DNA strands and using a guanine (guanine, G) base sequence close to AgNCs to enhance AgNCs A method for realizing specific fluorescence detection of HER2 based on the principle of fluorescence signals belongs to the technical field of biosensing. Background technique [0002] Human epidermal growth factor receptor 2 (HER2, also known as Neu or ErbB2) is a protein that promotes the growth of breast cancer cells. HER2 has become the most common marker of malignant tumors (D. J. Slamon, B. Leyland-Jones, S. Shak, H. Fuchs, V. Paton, V. N. Engl. J. Med. 2001, 344, 783-792.). The currently determined methods for detecting HER2 are: chromogenic in situ hybridization assay (CISH) (S.Kiyose, H.Igarashi, K.Nagura, T.Kamo, K.Kawane, H.Mori, T.Ozawa, M .Maeda, K.Konno, H.Hoshino, H.Konno, H.Ogu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6806G01N33/68G01N33/574
CPCC12Q1/6806G01N33/57415G01N33/6863G01N2333/71C12Q2563/137C12Q2563/107
Inventor 邓春艳张曼曼
Owner CENT SOUTH UNIV
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