A method and kit for the detection of adenosine triphosphate by a strand displacement signal amplification fluorescent sensor based on aptamers and carbon dots

A fluorescent sensor and adenosine triphosphate technology, applied in the field of analysis and detection, can solve the problems of complex operation, background signal interference, tedious and time-consuming, etc., and achieve the effect of simple experimental conditions, high selectivity detection, and improved sensitivity

Inactive Publication Date: 2021-02-19
NANJING MEDICAL UNIV
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Problems solved by technology

At present, there are many detection methods involving ATP, such as liquid chromatography-mass spectrometry, luciferase-mediated bioluminescence, gold nanoparticle-based colorimetry, etc., but these methods are complicated to operate, tedious and time-consuming, and susceptible to background Insufficient signal interference, etc.

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  • A method and kit for the detection of adenosine triphosphate by a strand displacement signal amplification fluorescent sensor based on aptamers and carbon dots
  • A method and kit for the detection of adenosine triphosphate by a strand displacement signal amplification fluorescent sensor based on aptamers and carbon dots
  • A method and kit for the detection of adenosine triphosphate by a strand displacement signal amplification fluorescent sensor based on aptamers and carbon dots

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Embodiment 1

[0050] a) Construct a fluorescent aptamer sensor: 2 μL aptamer strand (10 μM), 2 μL help strand (10 μM) and 20 μL CDs-signal strand (1 μM) were mixed evenly, and incubated at 37°C for 60 min; the above strands were pre-dissolved in 20 mM Tris -HCl;

[0051] b) Target circulating signal amplification: add 2 μL of 10 μM fuel strand and target ATP to the system, and react at room temperature for 110 min; fuel strand is pre-dissolved in 20 mM Tris-HCl;

[0052] c) Fluorescent detection of ATP: add graphene oxide to make the concentration 50 μg mL -1 , react at room temperature for 20 minutes, centrifuge, and put into a quartz cuvette for fluorescence detection;

[0053] d) Drawing of the standard curve: According to the above method, add different concentrations of ATP solutions to the system, the concentration is in the range of 0-800nM, incubate at room temperature, measure the respective fluorescence intensity values ​​F, and measure the blank fluorescence value F0 at the same...

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Abstract

The invention belongs to the technical field of analysis and detection, and relates to a method and a kit for detecting adenosine triphosphate by a chain displacement signal amplification fluorescence sensor based on an aptamer and carbon dots. The method comprises the following steps: a) hybridizing the carbon-dot-modified signal chain, the aptamer chain and the auxiliary chain to form a double-stranded probe; b) adding the reaction chain and the target ATP to the system of step a) to perform the first reaction; c ) adding graphene oxide, centrifuging after the second reaction, and performing fluorescence detection on the supernatant; d) according to the above method, adding different concentrations of ATP solutions to the system, measuring the respective fluorescence intensity values, and drawing the relative fluorescence intensity and ATP concentration Standard curve; e) According to the above method, add the test solution with unknown ATP concentration to the system, measure its fluorescence intensity value, and obtain the concentration of ATP in the test solution. The invention realizes the quantitative detection of ATP based on the change of the fluorescent signal, and has high specificity, high selectivity and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of analysis and detection, and more specifically relates to a method for detecting adenosine triphosphate by a chain displacement signal amplification fluorescent sensor based on aptamers and carbon dots, and a method for amplifying fluorescence by chain displacement signals based on aptamers and carbon dots A kit for the detection of adenosine triphosphate by a sensor. Background technique [0002] Adenosine triphosphate (ATP) is the main carrier of chemical energy in organisms and an important substrate for the enzyme activities of most organisms. It plays an important role in anti-arrhythmia, expansion of microvessels, and protection of cardiac muscle and nerve signals. The abnormal level of ATP in the body is closely related to many diseases, such as hypoxia, hypoglycemia, Parkinson's disease and some malignant tumors, etc. At the same time, the utilization efficiency of ATP is also an important indicato...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 周学敏罗洁萍沈心李昺之李晓芸
Owner NANJING MEDICAL UNIV
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