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Fluorescence biosensor based on light-up silver cluster probe and application thereof in miR-122 detection

A biosensor and probe technology, applied in the field of biosensors, can solve the problems of signal instability, complicated operation, poor reproducibility, etc., and achieve the effects of rapid response, low process cost and stable performance

Inactive Publication Date: 2019-12-13
UNIV OF JINAN
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Currently reported miR-122 detection techniques include electrochemical methods, colorimetric methods, PCR, surface plasmon resonance, etc. Some of these methods have problems such as complicated operation, high cost, unstable signal, and poor reproducibility

Method used

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  • Fluorescence biosensor based on light-up silver cluster probe and application thereof in miR-122 detection
  • Fluorescence biosensor based on light-up silver cluster probe and application thereof in miR-122 detection
  • Fluorescence biosensor based on light-up silver cluster probe and application thereof in miR-122 detection

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preparation example Construction

[0046] The preparation method of described biosensor comprises the following steps:

[0047] (1) Preparation of silver nanoclusters;

[0048] (2) The AgNC-SP / GrHP hybrid structure was formed by strand displacement reaction between the silver nanoclusters and the three-way structure in the homogeneous phase;

[0049] In the described preparation method, the preparation of silver nano clusters:

[0050] Silver nanoclusters were prepared according to the sodium borohydride reduction method reported in the literature. A mixed solution of DNA template strand AgNC-SP (15 μL, 100 μM) and silver nitrate (4.5 μL, 2 mM) for the synthesis of silver clusters was added to 76 μL of phosphate buffer (20 mM, pH 6.5) and reacted at 4 °C for 15 min. Afterwards, 4.5 μL of 2 mM sodium borohydride was added to the system, and left in the dark for more than 6 hours to form stable silver nanoclusters.

[0051] In the preparation method, a chain displacement reaction occurs between the silver nan...

Embodiment 1

[0055] In the described preparation method, the preparation of silver nano clusters:

[0056] Silver nanoclusters were prepared according to the sodium borohydride reduction method reported in the literature. A mixed solution of DNA template strand AgNC-SP (15 μL, 100 μM) and silver nitrate (4.5 μL, 2 mM) for the synthesis of silver clusters was added to 76 μL of phosphate buffer (20 mM, pH 6.5) and reacted at 4 °C for 15 min. Afterwards, 4.5 μL of 2 mM sodium borohydride was added to the system, and left in the dark for more than 6 hours to form stable silver nanoclusters.

[0057] Silver nanoclusters have been synthesized so far, and the main steps of the reaction process in the homogeneous solution are as follows:

[0058] Add IS strands (3 μL, 5 μM), GrHP probes (3 μL, 5 μM), BS strands (3 μL, 5 μM) and previously synthesized silver clusters (3 μL, 5 μM) into 1× buffer (50 mM NaCl, 10 mM Tris-acetic acid, 10 mM MgCl 2 , 100 μg·mL -1 BSA (pH7.9@25°C) was reacted at 37...

Embodiment 2

[0061] In the described preparation method, the preparation of silver nano clusters:

[0062] The nanoclusters were prepared according to the sodium borohydride reduction method reported in the literature. A mixed solution of DNA template strand AgNC-SP (15 μL, 100 μM) and silver nitrate (4.5 μL, 2 mM) for the synthesis of silver clusters was added to 76 μL of phosphate buffer (20 mM, pH 6.5) and reacted at 4 °C for 15 min. Afterwards, 4.5 μL of 2 mM sodium borohydride was added to the system, and left in the dark for more than 6 hours to form stable silver nanoclusters.

[0063] Silver nanoclusters have been synthesized so far, and the main steps of the reaction process in the homogeneous solution are as follows:

[0064] Add IS strands (3 μL, 5 μM), GrHP probes (3 μL, 5 μM), BS strands (3 μL, 5 μM) and previously synthesized silver clusters (3 μL, 5 μM) into 1× buffer (50 mM NaCl, 10 mM Tris-acetic acid, 10 mM MgCl 2 , 100 μg·mL -1 BSA (pH7.9@25℃) was reacted at 37℃ fo...

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Abstract

The invention relates to the technical field of biosensors, in particular to a fluorescence biosensor based on a light-up silver cluster probe and a preparation method of the fluorescence biosensor, and further relates to a G-rich base enhanced silver nano-cluster method. According to the detection mode provided by the invention, miR-122 detection is carried out through generation of a fluorescence signal, the hybridization of the miR-122 and the complementary strand thereof initiates the change of a three-path structure, a toehold end is exposed, and the silver cluster probe is crossed with GrHP clamp by virtue of the toehold end, so that a strand displacement reaction is initiated, the G-rich sequence is close to the silver cluster part, and the fluorescence intensity of the silver cluster is enhanced. The sensor has the advantages of being efficient, high in specificity, easy and convenient to operate, economical and free of marks, enzyme does not need to be used in a system, the defects and deficiencies of an existing detection method of miR-122 can be overcome, and rapid and accurate quantitative detection of miR-122 and early diagnosis of related diseases are achieved.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a fluorescent biosensor based on a light-up silver cluster probe and a preparation method thereof, and also to a method for enhancing silver nanoclusters with G-rich bases. Background technique [0002] MicroRNAs (MiRNAs) are a group of small non-coding RNAs that are expressed in various tissues and organs and participate in most physiological processes. Dysregulation of miRNAs contributes to several diseases, including cardiovascular disease, Parkinson's disease, and various cancers. Among many miRNAs, miR-122 is closely related to the occurrence of liver diseases. miR-122 is highly expressed in the liver of normal people, but its expression level is decreased in the liver of patients with liver cancer. In addition, miR-122 is also associated with chronic hepatitis C virus and drug-induced liver injury. Therefore, the sensitive detection of miR-122 is very important for the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6825C12Q1/6886C12N15/11
CPCC12Q1/6825C12Q1/6886C12Q2600/178C12Q2563/107C12Q2531/119
Inventor 王玉张雪刘素黄加栋宋晓蕾李莎莎王敬锋王海旺孙文玉王业茹
Owner UNIV OF JINAN
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