Method for detecting multiple targets based on nucleic acid aptamer fluorescence sensor

A fluorescent sensor and nucleic acid aptamer technology, applied in the field of analysis and detection, can solve the problems of low sensitivity, time-consuming detection, cumbersome steps, etc., and achieve the effect of low cost, strong versatility, and simple detection method

Inactive Publication Date: 2019-07-09
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This invention can overcome the disadvantages of low sensitivity, time-consuming detection, high cost and cumbersome steps of the existing analysis and detection methods

Method used

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  • Method for detecting multiple targets based on nucleic acid aptamer fluorescence sensor
  • Method for detecting multiple targets based on nucleic acid aptamer fluorescence sensor
  • Method for detecting multiple targets based on nucleic acid aptamer fluorescence sensor

Examples

Experimental program
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Effect test

Embodiment 1

[0041] A method for detecting target molecules such as interferon-γ, adenosine, and thrombin, comprising the following steps:

[0042] 1) Synthesis of DNA-AgNCs: Dissolve 1OD AgP1 template in 975 μL buffer 2, heat to 95°C, maintain for 10 min, then quickly transfer to an ice-water bath, rapidly cool down, then add 12 μL, 10 mM fresh silver nitrate solution, vortex After swirling and mixing, react at 4°C for 30 minutes in the dark, then quickly add 12 μL of 10 mM freshly prepared sodium borohydride solution, shake well and mix for 1 min, then react overnight at 4°C in the dark; dissolve 1OD of AgP2 template in 975 μL of buffer 2 , heated to 95°C, maintained for 10min, then quickly transferred to an ice-water bath, cooled rapidly, then added 16μL, 10mM freshly prepared silver nitrate solution, vortexed and mixed, and reacted at 4°C for 30min in the dark, then, quickly added 8μL, 10mM freshly prepared silver nitrate solution Sodium solution, shake and mix well for 1 min, then rea...

Embodiment 2

[0055] Selective experiment of detection method of the present invention

[0056] Repeat steps 1)-step 3) in Example 1, and replace the interferon-γ in step 3) with interfering substances such as bovine serum albumin, lysozyme, albumin, dopamine, L-cysteine, With other conditions unchanged, the fluorescence is detected, and the selectivity result diagram of the method for detecting the target interferon-γ is obtained, as shown in Figure 4 As shown, it can be seen from the figure that the method of the present invention has good selectivity to the target interferon-γ.

Embodiment 3

[0058] The recovery rate experiment of detection method of the present invention

[0059] Repeat step 1)-step 4) of 1) in the embodiment, in step 4), carry out adding sample detection respectively, add the adenosine of 7.8, 9.8, 11.8nM respectively, add the thrombin of 8, 10, 12nM or add 4.5, 6, 7.5nM interferon-γ was detected, and the recovery rate of this method in the actual sample detection was obtained. The results are shown in Table 2.

[0060] Table 2 Using this method to detect the recovery rate of adenosine, thrombin, and interferon-γ in serum samples

[0061]

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Abstract

The invention discloses a method for detecting multiple targets based on nucleic acid aptamer fluorescence sensor, which applies polypyrrole nanoparticles and DNA silver nanoclusters for the first time and establishes an aptamer sensing platform based on a steric hindrance effect.In addition, the invention obtains a good detection result based on the low non-specific adsorption characteristic ofthe polypyrrole nanoparticles, and realizes the detection of multiple targets based on the specific recognition characteristic of the aptamer sequence, improving the universal performance of the invention.The method for detecting multiple targets based on nucleic acid aptamer fluorescence sensor, provided by the invention, has the advantages of label-free property, no enzyme and high sensitivity,and can realize high-efficiency detection in biological samples.

Description

technical field [0001] The invention belongs to the technical field of analysis and detection, and more specifically relates to a method for multi-target detection based on a nucleic acid aptamer fluorescence sensor. Background technique [0002] In recent years, due to the emergence of the systematic evolution of ligands by exponential enrichment (SELEX) technology, nucleic acid aptamer sequences with high affinity to the target can be screened out from numerous nucleic acid sequences, and their specificity can be applied. Based on the principle of recognition, we can detect a variety of target molecules, ranging from small molecular amino acids to large molecular proteins. Using nucleic acid aptamers as biological recognition elements combined with signal conversion elements such as fluorescent groups, UV-responsive signal molecules, colorimetric sensing signal molecules, and electrochemical signal molecules can provide an ideal detection platform for the study of importan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6439
Inventor 周学敏洪俊丽王晶李昺之卢巧云李晓芸翁晨园严孝强
Owner NANJING MEDICAL UNIV
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