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DNA biosensor based on three-dimensional ordered gold doped nano titanium dioxide electrode as well as preparation method and application of DNA biosensor

A biosensor, three-dimensional ordered technology, applied in instruments, scientific instruments, electrochemical variables of materials, etc., can solve the problems of low detection sensitivity, high cost, and long time, and achieve simple solid loading conditions and solid loading capacity. The effect of large, good biocompatibility and conductivity

Inactive Publication Date: 2014-04-23
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In breast cancer detection methods, mammography is the most effective method, but for women under the age of 40, the detection sensitivity is relatively low due to the high density of breast cells; magnetic resonance imaging has good sensitivity in detecting breast cancer, but Expensive and time-consuming

Method used

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  • DNA biosensor based on three-dimensional ordered gold doped nano titanium dioxide electrode as well as preparation method and application of DNA biosensor
  • DNA biosensor based on three-dimensional ordered gold doped nano titanium dioxide electrode as well as preparation method and application of DNA biosensor
  • DNA biosensor based on three-dimensional ordered gold doped nano titanium dioxide electrode as well as preparation method and application of DNA biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Put 30mL absolute ethanol in a 100mL Erlenmeyer flask, add 0.3mL triethanolamine while stirring, then slowly add 0.7mL tetrabutyl titanate, continue stirring until the solution is uniform and transparent light yellow, then add 10mL HAuCl 4 , mixed solution of water and ethanol (0.5g / L of HAuCl 4 0.5mL, three times of water 3mL, absolute ethanol 6.5mL), stirring for 6h, the solution in the reaction will turn light yellow, slightly yellow, reddish, pink, until finally wine red, which is gold-doped nano-TiO 2 Sol. The experimental results of transmission electron microscopy show that the nanoparticles are uniformly distributed and the average particle size is less than 25nm.

[0047] The ITO electrode (50 mm×10 mm×1.2 mm) used was first soaked in lye for 30 min, then ultrasonicated in isopropanol, ethanol, and distilled water for half an hour, and then rinsed with secondary water for 3 times and allowed to dry naturally. Dry. Suspend the treated ITO in a weighing bottl...

Embodiment 2

[0049] The 3DOM GTD / ITO electrode prepared in Example 1 was rinsed with secondary water and dried at room temperature. 40 μL of 1.0 x 10 -5 mol / L 5'-Fc-ssDNA was dropped on the surface of 3DOM GTD / ITO electrode, and dried at 4 °C to prepare 5'-Fc-ssDNA / 3DOM GTD / ITO modified electrode, which was stored at 4 °C.

[0050] The UV-Vis absorption spectrum of the 5'-Fc-ssDNA / 3DOM GTD / ITO modified electrode has a DNA characteristic absorption peak at 260 nm, indicating that the DNA probe has been immobilized on the surface of the 3DOM GTD / ITO electrode. The peak position of DNA on the 5'-Fc-ssDNA / 3DOM GTD / ITO modified electrode was almost consistent with that of the free state, indicating that the DNA probe immobilized on the 3DOM GTD / ITO electrode still maintained its biological activity.

[0051] DNA probe molecules can generate obvious electrochemical signals on the surface of 3DOM GTD / ITO electrode, and the cyclic voltammetry curve of 5'-Fc-ssDNA / 3DOM GTD / ITO modified electrode s...

Embodiment 3

[0053] This example uses the principle of specific interaction between DNA probe (5'-Fc-ssDNA) and breast cancer gene fragment (ss1) two complementary DNA single strands, through the obvious change of DNA probe peak current before and after hybridization To detect ss1, to realize the electrochemical detection of breast cancer tumor genes.

[0054] The 5'-Fc-ssDNA / 3DOM GTD / ITO modified electrode was placed in 2 mL of ss1 solution, and hybridized at 37 °C for 1 h. Take out the electrode, wash it thoroughly with double distilled water to remove non-specifically adsorbed ss1, and dry it at 4 °C. Another random gene fragment ss2 was selected as the control group for the experiment, and the DNA sequence of ss2 was: 5'-GGT CAG GTG GGG GGT ACG CCA GG-3'.

[0055] The electrochemical detection adopts a three-electrode system, with the prepared 5'-Fc-ssDNA / 3DOM GTD / ITO modified electrode and the electrode after hybridization as the working electrode, the saturated calomel electrode as ...

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Abstract

The invention discloses a DNA biosensor based on a three-dimensional ordered gold doped nano titanium dioxide electrode and a preparation method as well as a method for detecting breast cancer genes by using the sensor. The preparation method comprises the following steps: preparing a three-dimensional ordered gold doped nano titanium dioxide film electrode (3DOMGTD / ITO) on an ITO (indium tin oxide) electrode by adopting a template method; then dropping a DNA probe (5'-Fc-ssDNA) labeled with ferrocene at 5' end on the electrode, thereby preparing a modified electrode, namely the DNA biosensor; placing the modified electrode in a solution of a breast cancer gene segment, hybridizing, detecting the variation of hybridization signals by adopting a differential pulse volt-ampere method, and carrying out electrochemical detection on the breast cancer gene. According to the invention, the label-type DNA biosensor is prepared by utilizing the specificity recognition effect between the DNA probe and the breast cancer gene, and the DNA biosensor has favorable stability, reproducibility and electrode regeneration, and can be applied to the field of DNA hybridization and tumor electrochemical detection.

Description

technical field [0001] The present invention relates to a DNA biosensor and its preparation and application, in particular to a labeled DNA biosensor based on a three-dimensional ordered nano-titanium dioxide electrode and its preparation, and the application of the biosensor for breast cancer tumor gene detection. The invention belongs to the technical field of DNA biosensor and the technical field of bioelectrochemistry. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, seriously affecting women's physical and mental health and even threatening their lives. Studies have found that the variation of base sequences in DNA molecules is related to tumors and many human hereditary diseases. Therefore, research uses specific DNA probes to identify breast cancer genes and develop fast, simple and reliable detection methods for breast cancer genes. , It is of great significance to provide evidence for early diagnosis and treatment of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/26
Inventor 常颖萃杜江燕
Owner NANJING NORMAL UNIVERSITY
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