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Method for detecting 17bata-estradiol by using resonance scattering spectrometry based on functional nucleic acids

A resonance scattering, functional nucleic acid technology, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of prone to false positives, long detection time, high cost, and achieve the effect of significant technological progress, low cost, and good stability.

Inactive Publication Date: 2015-06-03
SHANGHAI APPLIED TECHNOLOGIES COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above-mentioned technical problems in the prior art, the present invention provides a method for detecting 17β-estradiol based on functional nucleic acid using resonance scattering spectroscopy, which solves the problem of detecting 17β-estradiol in the prior art. The method is prone to false positives, high cost, and technical problems such as long detection time

Method used

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  • Method for detecting 17bata-estradiol by using resonance scattering spectrometry based on functional nucleic acids
  • Method for detecting 17bata-estradiol by using resonance scattering spectrometry based on functional nucleic acids
  • Method for detecting 17bata-estradiol by using resonance scattering spectrometry based on functional nucleic acids

Examples

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Effect test

Embodiment 1

[0028] Example 1 Sodium chloride concentration in the optimized system

[0029] In a 1.5mL centrifuge tube, add 100μL of nano-gold solution, then add propanesulfonic acid buffer (concentration is 10mmol / L, pH is 7.0), mix well, and sequentially add sodium chloride solution with a concentration range of 0-160mmol / L, After five minutes, the resonance scattering value was measured, the relative resonance scattering value was obtained, and a curve was drawn. The sodium chloride concentration corresponding to the highest peak was 120 mmol / L, such as figure 2 shown.

Embodiment 2

[0030] Example 2 Optimizing the concentration of 17β-estradiol aptamer in the system

[0031] The 17β-estradiol aptamer was purchased from Shanghai Shenggong. The received sample was prepared into a 1 μmol / L mother solution, and then the aptamer with a concentration range of 0-40 nmol / L was added to 100 μL of nano-gold solution. For aptamer-gold nanoparticles, after incubation for 15 minutes, add a certain concentration of estradiol solution, incubate for 15 minutes, add the optimized sodium chloride solution, adjust the volume to 500 μL with propanesulfonic acid buffer, and measure the resonance scattering value after five minutes , draw a curve, the aptamer concentration corresponding to the highest peak is 20nmol / L, such as image 3 shown.

Embodiment 3

[0032] Embodiment 3 The process of using the method of the present invention to detect whether the milk sample contains 17β-estradiol specifically includes the following steps:

[0033](1) Prepare a detection system with known 17β-estradiol concentration: take 11 centrifuge tubes to which 10 μL and 1 μmol / L 17β-estradiol aptamer were added respectively, add 100 μL nano-gold solution, mix well and place the centrifuge tubes into the centrifuge tubes. Incubate at 25°C for 15min, and then add 10μL of 17β-estradiol solutions of different concentrations respectively, so that the content of 17β-estradiol in the entire detection system is maintained at 0.1-3.0μmol / L, and then fully mixed before Incubate the centrifuge tube at 25°C for 15 min, add 30 μL, 2 mol / L sodium chloride solution to the above mixture, mix well, and adjust the buffer volume to 500 μL, which is reserved for the following determinations;

[0034] (2) Take another centrifuge tube and replace the 17β-estradiol solut...

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Abstract

The invention discloses a method for detecting 17bata-estradiol by using resonance scattering spectrometry based on functional nucleic acids. The method comprises the following steps: preparing an aptamer-nanogold probe sequence, wherein the aptamer is a section of functional nucleotide sequence, and the nucleotide sequence is 5'-GCT-TCC-AGC-TTA-TTG-AAT-TAC-ACG-CAG-AGG-GTA-GCG-GCT-CTG-CGC-ATT-CAA-TTG-CTG-CGC-GCT-GAA-GCG-CGG-AAG-C-3'; combining nanogold with the aptamer to form a stable aptamer-nanogold probe; and adding a to-be-detected aqueous solution which is suspected to contain 17bata-estradiol to a probe solution, performing incubation, adding a sodium chloride solution, and judging the content of 17bata-estradiol in the to-be-detected solution by measuring the strength of a resonance scattering signal. According to the method disclosed by the invention, the detection of 17bata-estradiol can be achieved by comparing the changes of resonance scattering values.

Description

technical field [0001] The present invention in the field of food relates to a method for detecting food, in particular to a method for detecting 17β-estradiol, specifically a method for detecting 17β-estradiol based on functional nucleic acid using resonance scattering spectroscopy. Background technique [0002] Environmental estrogens, also known as "endocrine disrupting compounds", are a class of pollutants that are widely present in the environment, which can disrupt the normal endocrine function of the human and animal bodies, and change the body's intracellular signaling processes during development and adulthood, thereby causing Their reproductive, immune, nervous and other systems of a variety of lesions. However, 17β-estradiol is one of the most powerful environmental estrogens, and it is commonly found in various environments, especially in water environments, such as milk, honey, etc. 17β-estradiol is synthesized in animals and humans, and has the activity of end...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 夏兵王鲁梅叶璟张东伟杜玉萍邱志浩
Owner SHANGHAI APPLIED TECHNOLOGIES COLLEGE
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