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Method for detecting lead ions based on aptamer DNAzyme

A technology of aptamers and substrate chains, which can be used in material analysis by observing the influence of chemical indicators, measurement of color/spectral characteristics, analysis by making materials undergo chemical reactions, etc., which can solve the problem of expensive equipment and discomfort Universal application of scale, limited scope of use of methods, etc.

Inactive Publication Date: 2018-03-06
杨蕾
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Problems solved by technology

[0005] Technical problem to be solved: Due to the detection of Pb 2+ The instrument method is expensive, and the sample requires professional operators to carry out complicated pretreatment processes, so it is not suitable for large-scale general application, which limits the scope of use of this method

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  • Method for detecting lead ions based on aptamer DNAzyme

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Embodiment 1

[0026] An aptamer-based DNAzyme for Pb 2+ The detection method comprises the steps of:

[0027] (1) Biotin-modified Pb 2+ DNAzyme substrate strand coated microtiter plates

[0028] Firstly, streptavidin was dissolved with 0.01M carbonate buffer solution of pH 9.6, and then diluted with carbonate buffer solution to a coating concentration of 1.0ng / mL, with a coating volume of 100μL / well Coated onto 96-well ELISA plate and incubated in an oven at 37°C for 2 hours. After incubation, the ELISA plate was washed 3 times with 0.01M PBS washing solution of pH 7.4 containing 0.5% Tween20, and finally patted dry; biological prime-modified Pb 2+ The substrate strand of the DNAzyme was dissolved in 0.01M PBS with pH 7.4, one end of the DNAzyme substrate strand was connected with G-quadruplex DNA, and then diluted to a concentration of 4.0ng / mL, and added to the streptavidin package Add 100 μL to each well of the well-prepared ELISA plate, incubate it in a 37°C oven for 1 hour, take it...

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Abstract

The invention relates to a method for detecting Pb<2+> based on aptamer DNAzyme and belongs to the field of biological detection. The method provided by the invention comprises the following steps: coating an enzyme-labeled board by using a substrate chain of the DNAzyme of the biotin-modified Pb<2+>, hybridizing a DNAzyme chain of the Pb<2+> and the substrate chain to form a double-chain structure, detecting the Pb<2+> and determining an UV absorption value. The method provided by the invention has the benefits that by means of a catalytic effect of the Pb<2+>-dependent DNAzyme and an enzymecatalytic effect of a G quadruple-hemin compound, in the presence of different concentrations of Pb<2+>, the color development difference of a G quadruple-hemin compound-catalyzed substrate is produced; according to a corresponding relation between the Pb<2+> concentration and the UV absorption value, the detection on the Pb<2+> is realized. The method provided by the invention is high in sensitivity, low in detection limit, good in specificity, quick in detection speed and low in cost, thereby being an ideal Pb<2+> detection method.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to a method based on aptamer DNAzyme for Pb 2+ method of detection. Background technique [0002] In recent years, with the increasing environmental pollution caused by heavy metals, heavy metal pollution has become a growing ecological problem, and has also become a public health issue of global concern, and lead and its compounds are one of the heavy metal pollution important pollutants. There is a large amount of lead in the fields of automobile exhaust, coal product combustion flue gas, industrial electroplating, metallurgical wastewater, lead-acid batteries, etc. At the same time, beverages, inferior foods, canned foods and other foods may also contain heavy metals. Lead cannot be degraded in water. It has serious damage to the nervous system, bone hematopoietic function, digestive system, and respiratory system of the human body, especially children whose brains are in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/33
CPCG01N21/33G01N21/78
Inventor 杨蕾
Owner 杨蕾