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17 beta-estradiol colorimetric detecting method based on nanogold collected by surfactant

A surfactant and estradiol technology, applied in the field of 17β-estradiol detection, to achieve the effect of good selectivity, fast detection method and high sensitivity

Inactive Publication Date: 2015-11-11
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the aptamer can also be combined with the cationic surfactant CTAB to form a supramolecular structure. The present invention is based on the nucleic acid aptamer, utilizes the cationic surfactant CTAB to gather nano-gold, and uses a multifunctional microplate reader to measure the absorbance to realize the detection of 17β -quantitative detection of estradiol, the method constructed by the present invention has not been reported in the prior art

Method used

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  • 17 beta-estradiol colorimetric detecting method based on nanogold collected by surfactant
  • 17 beta-estradiol colorimetric detecting method based on nanogold collected by surfactant
  • 17 beta-estradiol colorimetric detecting method based on nanogold collected by surfactant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Optimization of CTAB Concentration of Cationic Surfactant

[0035] Add 200μL of nano-gold solution to a 1.5mL centrifuge tube, then add a certain amount of PBS buffer (concentration of 10mmol / L, pH of 7.0), mix well, and sequentially add surfactants with a concentration range of 0.4~1.8μmol / L CTAB solution, make the total volume of the system reach 500μL, then take 200μL of the reaction solution in a 96-well plate, use a multi-functional microplate reader to measure the absorbance at 520nm and 650nm, calculate the ratio A=A650 / A520, draw the curve, the peak value corresponds to the highest The concentration of surfactant CTAB is 1.4μmol / L, such as figure 2 Shown.

Embodiment 2

[0036] Example 2. Optimization of functional nucleic acid aptamer concentration

[0037] (1) The aptamer was purchased from Shanghai Shenggong, the received samples were prepared into 100nmol / L mother liquor, and then the aptamer with a concentration range of 1~6nmol / L was added to 500nmol / L of 17β-estradiol. Incubate in the solution for 20 minutes, add the optimized surfactant CTAB solution in Example 1 and a certain amount of PBS buffer, mix thoroughly, then place the centrifuge tube at 25°C and incubate for 20 minutes, and add 200 μL to the above mixture Nano-gold solution, mix thoroughly so that the total volume of the detection system solution reaches 500μL, and reserve it for the following measurements;

[0038] (2) Set up another set of aptamer solutions with a concentration range of 1~6nmol / L, and replace the 17β-estradiol solution with ultrapure water to make the total volume of the detection system reach 500μL. Follow the steps in step (1) above Used as a blank control ...

Embodiment 3

[0040] Example 3. Specificity experiment

[0041] (1) Take 10 centrifuge tubes each with 15μL, 100nmol / L aptamer, and then add 25μL, 10μmol / L 17β-estradiol, estriol, bisphenol A, progesterone, ethylene estrol, DDT, tetracycline, aminocephalosporanic acid, copper ion solution, cadmium ion solution, and then mix well, then place the centrifuge tube at 25℃ and incubate for 20min, add 14μL, 50μmol / L surfactant to the above mixture The CTAB solution and a certain amount of PBS buffer are mixed thoroughly and incubated at 25°C for 20 minutes. Add 200μL of nano-gold solution to the above mixture and mix thoroughly to make the total volume of the detection system solution reach 500μL, and reserve for the following determination use;

[0042] (2) Take another centrifuge tube and add 25μL of ultrapure water, treat it according to the above step (1) as a blank control system solution;

[0043] (3) Take 200μL of the above two groups of reaction solutions in a 96-well plate, use a multifunctio...

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Abstract

The invention discloses a 17 beta-estradiol colorimetric detecting method based on nanogold collected by a surfactant. The method comprises the following steps: adding a 17 beta-estradiol sample to be detected into an aptamer solution, uniformly mixing and incubating to obtain a mixed solution A; adding the surfactant and a buffer solution into the mixed solution A, uniformly mixing and incubating to obtain a mixed solution B; adding a nanogold solution into the mixed solution B and mixing to obtain a solution C; determining that the light absorption value ratio A650 / 520 of a blank control solution and the solution C at the positions of 650 nm and 520 nm are respectively A0 and A, and obtaining delta A according to the formula that delta A=A-A0; determining the content of 17beta-estradiol corresponding to delta A according to a standard curve of the content of 17beta-estradiol. The provided detecting method is quick, cheap, high in sensitivity and good in selectivity, the minimum detection limit is 1.96 nmol / L, and the defects of consuming time, relying on large-scale instruments and equipment, being high in cost, and the like, in the conventional detecting method are overcome.

Description

Technical field [0001] The present invention belongs to the detection field of 17β-estradiol in water environment. It is a 17β-estradiol colorimetric detection method based on surfactant aggregated nano-gold, and particularly relates to a functional nucleic acid aptamer using cationic surfactant A method for colorimetric detection of 17β-estradiol by aggregated nano-gold. Background technique [0002] 17β-estradiol is a natural estrogen, and it is widely present in the natural environment. It can enter the human body from the external environment, thereby disrupting the function of the human endocrine system and causing the human body to be in the nervous system, immune system and reproductive development systems. Of lesions. Therefore, it is of great practical significance to develop a simple, rapid, and sensitive detection method for 17β-estradiol. [0003] The traditional detection methods of 17β-estradiol include high performance liquid chromatography, gas chromatography-mass...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/5308
Inventor 王鲁梅夏兵张东伟詹深山吕晶杜高尚徐涵楚
Owner SHANGHAI JIAO TONG UNIV
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