Preparation and application of aptamer colorimetric sensor for polychlorinated biphenyl 77 detection

A nucleic acid aptamer and colorimetric sensor technology, which is applied in the preparation and application of nucleic acid aptamer colorimetric sensors, can solve the problems of complicated operation and expensive instruments, and achieve simple preparation, low synthesis cost, high sensitivity and The effect of highly selective analytical detection

Inactive Publication Date: 2017-09-12
TONGJI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide an analysis method based on a certain base sequence for a nucleic acid-adapted colorimetric sensor in view of the existing methods for detecting PCB 77, which are expensive, complicated to operate, and time-consuming to analyze.

Method used

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  • Preparation and application of aptamer colorimetric sensor for polychlorinated biphenyl 77 detection

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Experimental program
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Effect test

Embodiment 1

[0019] A nucleic acid aptamer colorimetric sensor based on a certain base sequence for rapid detection of PCB 77. The gold nanoparticle sol is used as the matrix. PCB 77 competes with gold nanoparticles to bind to the nucleic acid aptamer to cause the aggregation of gold nanoparticles. Changes to realize the analysis of PCB 77. The specific preparation steps of the nucleic acid aptamer colorimetric sensor are as follows:

[0020] (1) All glass instruments are soaked in aqua regia, cleaned with high-purity water and dried for later use. During preparation, add 100 mL of chloroauric acid (HAuCl) with a concentration of 0.05 mol / L to a clean beaker 4 ) Solution, continue to heat to boiling; under continuous stirring, add 10ml of 0.0388mol / L sodium citrate solution to the boiling HAuCl 4 In the solution, the heating was stopped after the reaction continued for 20 minutes. After the solution is cooled to room temperature, it is filtered with a 0.22 μm filter membrane to obtain a gold...

Embodiment 2

[0025] For the two concentration parameters and reaction time parameters in the preparation process of nucleic acid aptamer colorimetric sensors: the optimal concentration of nucleic acid aptamer and sodium chloride, and the optimal amount of reaction time were determined. Specifically: take a series of gold nanoparticle sols, add different concentrations of aptamer solution and sodium chloride solution to them, and prepare the nucleic acid aptamer at a concentration of 1.40×10 -8 ~1.40×10 -7 A series of nucleic acid aptamer colorimetric sensor solutions in the range of mol / L and the concentration of sodium chloride in the range of 0.01~0.04mol / L. Measure the A of the above series of solutions with UV-Vis spectrophotometer respectively 646 / A 520 . The research results show that when the concentration of aptamer is 3.8×10 -7 mol / L, when the concentration of sodium chloride is 0.0345mol / L, A 646 / A 520 The ratio is the largest. Therefore, the optimal concentrations of aptamer an...

Embodiment 3

[0027] Take PCB 77 liquid sample and prepare it with n-hexane to obtain 1.0×10 -3 mol / L PCB 77 mother liquid was diluted with different volumes of ethanol to prepare a series of PCB 77 standard solutions with different concentrations. Take the nucleic acid aptamer colorimetric sensor solution prepared in Example 1, respectively add PCB77 solutions of different concentrations into it, and after acting at 25° C. for 3 minutes, the ultraviolet-visible absorption spectrum of the solution is measured with an ultraviolet-visible spectrophotometer. The research results show that with the increase of PCB 77 concentration in the test system, the characteristic plasmon resonance absorption peak of gold nanoparticles is red-shifted from 520nm to 646nm. The higher the concentration of PCB 77, the absorbance value A of gold nanoparticles at 646nm 646 The bigger, the absorbance value A at 520nm 520 The smaller the ratio A 646 / A 520 Bigger. Use A 646 / A 520 The linear relationship with the l...

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Abstract

The invention relates to preparation and application of an aptamer colorimetric sensor for polychlorinated biphenyl 77 detection. An aptamer with a certain base sequence and specific PCB (polychlorinated biphenyl) 77 recognition capability and the colorimetric analysis technology are combined for constructing the PCB 77 aptamer colorimetric sensor. Due to adoption of the specific PCB 77 recognition aptamer as a recognition element, sensitivity and selectivity of the colorimetric sensor in PCB 77 detection are greatly improved. A PCB 77 detection linear range is 5*10<-10>-9*10<-7>mol/L, and a detection limit reaches 5*10<-11>mol/L. Compared with the prior art, the method has advantages of simplicity and convenience in operation, quickness in response, easiness in online detection and the like.

Description

Technical field [0001] The invention belongs to the field of water pollution detection and treatment, and in particular relates to the preparation and application of a nucleic acid aptamer colorimetric sensor for detecting polychlorinated biphenyl 77. Background technique [0002] Polychlorinated biphenyl 77 (PCB 77) is a type of persistent organic environmental pollutant with high toxicity and low concentration that is widely present in the environment. Since the 20th century, PCB pollution incidents have occurred in many parts of the world, causing irreparable impacts on human and animal health and the environment. Compared with other pollutants, it has the characteristics of high toxicity and low lethal dose. PCBs are found in many sewage treatment plants, along the rivers and estuaries, and they will exist in water bodies, and will remain in the soil. Finally, they will be enriched in humans or animals through the food chain, which will affect the nerves and reproduction of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N1/34
CPCG01N21/314G01N1/34G01N2021/3155
Inventor 赵国华程若洁
Owner TONGJI UNIV
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