Development and application of Ag nano-cluster electrochemiluminescence sensor based on in-situ synthesis

An electrochemistry, in-situ synthesis technology, applied in the direction of chemiluminescence/bioluminescence, analysis by chemical reaction of materials, etc., to achieve the effect of high selectivity, improved sensitivity and excellent accuracy

Inactive Publication Date: 2018-06-29
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the combination of DNase cycle amplification technology and nano-amplification effect, and applied to highly sensitive ECL biosensing analysis.

Method used

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  • Development and application of Ag nano-cluster electrochemiluminescence sensor based on in-situ synthesis
  • Development and application of Ag nano-cluster electrochemiluminescence sensor based on in-situ synthesis
  • Development and application of Ag nano-cluster electrochemiluminescence sensor based on in-situ synthesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1. Preparation of electrochemiluminescence biosensor and detection of thrombin

[0026] DNase cycle shearing process: First, 1μM hairpin DNA was preactivated at 37°C for 1h, then 10μL of thrombin (2μM) of different concentrations was added and incubated in Tris-HCl buffer at 37°C for 1h. Subsequently, the shearing and circulation process was started, and the substrate (5 μL, 3 μM) and 10 μL of 0.01M zinc acetate solution were added to the above buffer solution, and finally the volume of the above solution was 50 μL and incubated at 37°C for 50 minutes. Recorded as solution s1.

[0027] Before the DNA is diluted, add the corresponding amount of buffer solution to the tube according to the instructions, and the concentration is 10 -4 mol / L DNA solution, store it at 4℃ for later use.

[0028] Electrode treatment: first, use 1.0, 0.3, 0.05μm α-Al for gold electrode 2 O 3 The powder is polished for 5 minutes, then rinsed thoroughly with ultrapure water and dried with nitro...

Embodiment 2

[0030] Example 2. Preparation of electrochemiluminescence biosensor and detection of thrombin

[0031] Change "Then start the shearing and cycling process, add the substrate (5μL, 3μM) and 10μL 0.01M zinc acetate solution to the above buffer, and finally make the volume of the above solution 50μL, and incubate at 37°C for 50min." "Then start the shearing and cycling process, add the substrate (5μL, 3μM) and 10μL 0.01M zinc acetate solution to the above buffer, finally make the volume of the above solution 50μL, and incubate at 37°C for 60min." Other prepared The conditions were the same as in Example 1, and a biosensor with similar morphology and properties to Example 1 was obtained. The results of thrombin detection are the same as in Example 1.

Embodiment 3

[0032] Example 3. Preparation of electrochemiluminescence biosensor and detection of thrombin

[0033] Change "Next take 8μL of SH-DNA (0.5μM) diluted with fixation buffer (10mM Tris-HCl, 1mM EDTA, 10mM TCEP, 0.1M NaClpH=7.4) and drop it on the electrode surface" to "Next take 8μL SH-DNA (1μM) diluted with fixation buffer (10mM Tris-HCl, 1mM EDTA, 10mM TCEP, 0.1M NaCl pH=7.4) was dripped onto the electrode surface". The other preparation conditions are the same as in Example 1, and a biosensor with morphology and properties similar to that in Example 1 is obtained. The results of thrombin detection are the same as in Example 1.

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Abstract

The invention discloses an application of an electrochemiluminescence biosensor based on cytosine (C)-containing cyclic DNA sequences where in-situ reduced Ag nano-clusters are enriched as signal probes and adopting dual amplification strategy to detection of target thrombin. According to the technical scheme, DNA cleavage enzyme with hairpin DNA recognition capability and catalysis effect is designed, when target thrombin exists, hairpin DNA is opened, and substrate DNA is cleaved under the action of Zn<2+>; a large quantity of cytosine (C)-containing cyclic DNA sequences are aggregated through HCR (hybridization chain reaction), in-situ reduction of AgNO3 is performed on the electrode surface by NaBH4, a large quantity of Ag nano-clusters are formed, and the electrochemiluminescence biosensor with dual amplification effect is prepared. The sensor is subjected to luminescence detection, and a linear relation is formed between light-emitting signals and concentration of a to-be-testedsample. The Ag nano-cluster signal probes and the DNA dual amplification technology are combined for rapid and high-sensitivity detection of thrombin, and the electrochemiluminescence biosensor has great application potential in early clinical analysis and detection.

Description

Technical field: [0001] The invention relates to a new method for the development of a silver nano-cluster electrochemiluminescence biosensor based on in-situ synthesis; and an analysis application of the electrochemiluminescence biosensor combined with dual signal amplification technology to detect thrombin. Background technique: [0002] Tracking, analyzing and detecting the biological activity of biological molecules such as DNA and proteins are closely related to human health [Fields, S. Science 2001, 291, 1221-1224.]. However, the concentration of biomolecules in the process of disease diagnosis and in vivo exploration is very low, and more sensitive methods are still needed to detect these biomolecules. Electrochemiluminescence (ECL) analysis has low background and high sensitivity, and has been widely used in biological analysis. Nanoparticles are widely used in the field of biomedicine due to their potential functions in diagnosis and treatment. [Dreaden, E.C.; Alkilany...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
CPCG01N21/76
Inventor 接贵芬葛君君
Owner QINGDAO UNIV OF SCI & TECH
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