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Electrode modified based on graphene oxide, gold nanotube and locked nucleic acid probe and its preparation method and application

A technology for locking nucleic acid probes and gold nanotubes, applied in the field of electrode preparation, can solve the problems of DNA probes with low specificity, weak specificity, base mismatch, etc.

Active Publication Date: 2017-01-04
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Specificity and sensitivity are important indicators for evaluating the detection performance of sensors. Most of the sensors reported in current studies are immobilized with DNA probes, and their disadvantages are low sensitivity and low specificity.
The specificity of DNA probes is relatively low due to the weak binding force of DNA probes when hybridizing to longer target sequences, often resulting in base mismatches

Method used

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  • Electrode modified based on graphene oxide, gold nanotube and locked nucleic acid probe and its preparation method and application
  • Electrode modified based on graphene oxide, gold nanotube and locked nucleic acid probe and its preparation method and application
  • Electrode modified based on graphene oxide, gold nanotube and locked nucleic acid probe and its preparation method and application

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Experimental program
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Embodiment 1

[0031] The preparation method based on the electrode modified by graphene oxide, gold nanotube and locked nucleic acid probe comprises the following steps of graphene oxide, gold nanotube and locked nucleic acid probe modified gold electrode and preparation of signal amplification probe:

[0032] (1) Preparation of graphene oxide ZnO sol-gel composite: Weigh 0.001g graphene oxide in a small beaker, add 1mL of ultrapure water, and then place it in an ultrasonic instrument to prepare a graphene oxide suspension, Take 90 μL of graphene oxide suspension, 30 μL of ZnO sol-gel and 60 μL of absolute ethanol and mix well to prepare a graphene oxide ZnO sol-gel mixture, and store it in the dark at 4°C for later use;

[0033] (2) Pretreated electrodes: use 0.3μm and 0.05μm Al for the electrodes 2 o 3 The powder is ground and polished to a mirror surface, rinsed with ultra-pure water after each grinding, and then ultrasonically washed in nitric acid, acetone and ultra-pure water for 5 m...

Embodiment 2

[0036] Embodiment 2, set up electrochemical biosensor

[0037] The electrode modified by graphene oxide, gold nanotube and locked nucleic acid probe is used as the working electrode, the saturated calomel electrode is used as the reference electrode, and the platinum electrode is used as the counter electrode to form an electrochemical biosensor. Connect the assembled electrochemical biosensor to the electrochemical workstation, and in pH7.4 containing 2mmol / L K 3 Fe(CN) 6 / K 4 Fe(CN) 6 Cyclic voltammetry scanning was carried out in PBS with 5mmol KCl, the working potential was -0.3V~0.7V, and the scanning speed was 50mV / s.

[0038] Hybridization reaction: immerse the gold electrode modified by graphene oxide, gold nanotube and locked nucleic acid probe into 150 μl of different concentrations of NDM-1 target sequence solution (10 -6 μg / L, 10 -5 μg / L, 10 -4 μg / L, 10 -3 μg / L, 10 -2 μg / L, 10 -1 μg / L, 1.0μg / L, 10 1 , 10 2 μg / L), and then add the signal amplification pro...

Embodiment 3

[0040] Embodiment 3, step-by-step characterization based on graphene oxide, gold nanotubes and locked nucleic acid probes

[0041] Use cyclic voltammetry to characterize the electrochemical characteristics of the electrode during the modification and detection process. The specific detection method is: immerse the modified electrode in 150 μL of NDM-1 target sequence solution with a concentration of 100 μg / L, and then add lock to the reaction system. Nucleic acid probe solution, hybridized at a constant temperature of 37°C for 45 minutes. During the hybridization process, the sensor used a three-electrode system to measure the current change, and obtained a concentration of 2mmol / L K at pH 7.4 for different electrodes. 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ] and the cyclic voltammetry curve (CV) figure in the PBS solution of 5mmol / l KCl, as figure 1 shown.

[0042] figure 1 Among them, curve a is graphene oxide ZnO sol-gel and gold nanotube modified electrode containing 2mmol / LK at...

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Abstract

The invention discloses an electrode modified based on graphene oxide, gold nanotubes and locked nucleic acid probes and its preparation method and application. The electrode is a gold electrode modified by graphene oxide, gold nanotubes and locked nucleic acid probes and signal amplification Probe composition, with this electrode as working electrode, platinum electrode as counter electrode, saturated calomel electrode as reference electrode, the PBS solution containing 2mmol / L K3Fe(CN)6-K4Fe(CN)6 and 5mmol / L KCl is The test base solution is used to build an electrochemical biosensor, which can quickly, specifically and sensitively detect the NDM-1 gene, and provide a simple, fast, sensitive and specific tool for the detection of multi-drug resistant bacteria, which has a good market value.

Description

technical field [0001] The invention belongs to the field of electrochemical detection, and in particular relates to an electrode modified based on graphene oxide, gold nanotube and locked nucleic acid (Locked nucleic acid, LNA) probe, and also relates to a preparation method and application of the electrode. Background technique [0002] NDM-1 multidrug-resistant bacteria refer to bacteria carrying NDM-1 drug-resistant genes, which belong to Gram-negative drug-resistant bacteria. The full name of NDM-1 is "New Delhi metallo-β-lactamase", which is a highly efficient drug-resistant enzyme. Studies have shown that multidrug-resistant bacteria are spreading rapidly around the world, and NDM-1 has been found in Shigella, an important human pathogen. Bacteria carrying the NDM-1 drug resistance gene can be transmitted through drinking water and other channels, and the symptoms are intestinal infection. This type of bacteria is resistant to almost all antibiotics, and the death ra...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48
Inventor 王云霞张立群府伟灵
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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