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Toxin detection method based on G-quadruplex nuclease

A detection method, a quadruplex technology, applied in the field of immunoanalytical chemistry, can solve problems such as the detection of microcystins that have not been seen

Inactive Publication Date: 2013-09-11
CHANGSHU INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been used for the detection of microcystin (MC-LR)

Method used

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  • Toxin detection method based on G-quadruplex nuclease
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  • Toxin detection method based on G-quadruplex nuclease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Synthesis of gold nanoparticles;

[0040] Gold nanoparticles with a diameter of 25 nm were synthesized by referring to Frens' trisodium citrate reduction method. The specific operation steps are as follows: Add 50 mL of the prepared 1.0 mM chloroauric acid solution into the Erlenmeyer flask, place it on a constant temperature electromagnetic stirrer and heat it to boiling for 5 minutes, then quickly add 0.6 mL, 38.8 mM citric acid Continue to stir and heat the trisodium solution for 6 minutes. The color of the solution changes from light yellow to wine red. At this time, gold nanoparticles are formed. Continue to heat for 10 minutes and then stop. Continue to stir to cool it down, and store it at 4°C for later use.

[0041] (2) Functionalization of gold nanoparticles;

[0042] The functionalization of gold nanoparticles refers to modifying DNA to gold nanoparticles through the sulfhydryl group of nucleic acid and the gold sulfhydryl bond on the surface of gold part...

Embodiment 2

[0058] Compared with Example 1, the only difference is that step 1 is different, specifically:

[0059] Add 50 mL of the prepared 1.0 mM chloroauric acid solution into the Erlenmeyer flask, place it on a constant temperature electromagnetic stirrer and heat it to boiling for 5 minutes, then quickly add 0.6 mL, 38.8 mM trisodium citrate solution, and continue stirring Heating for 4 minutes, the color of the solution changed from light yellow to wine red, at this time gold nanoparticles were formed, stop heating after 10 minutes, continue stirring to cool down, and store at 4°C for later use.

Embodiment 3

[0061] Compared with Example 1, the only difference is that step 1 is different, specifically:

[0062] Add 50 mL of the prepared 1.0 mM chloroauric acid solution into the Erlenmeyer flask, place it on a constant temperature electromagnetic stirrer and heat it to boiling for 10 minutes, then quickly add 0.6 mL, 38.8 mM trisodium citrate solution, and continue stirring Heating for 6 minutes, the color of the solution changed from light yellow to wine red, at this time gold nanoparticles were formed, stop heating after 15 minutes, continue stirring to cool down, and store at 4°C for later use.

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Abstract

The invention discloses an MC-LR (microcystin-LR) detection method based on G-quadruplex nuclease, and belongs to the technical field of immunoassay chemistry. A quadruplex-hemin compound with similar peroxidase activity is successfully prepared and coupled to a gold nanoparticle surface, signals can be amplified, and finally, a gold nanoparticle-quadruplex conjugate is successfully marked on an antibody. By the aid of the novel antibody marker, a novel immunodetection method based on G-quadruplex DNA (deoxyribonucleic acid) is used for detecting microcystins. The method is simple, rapid and convenient, fine sensitivity and selectivity are embodied, detection sensitivity is improved by the signal amplification function of quadruplex, the microcystins in water can be simply, rapidly and highly sensitively detected, and international advanced level is reached.

Description

technical field [0001] The invention relates to a new method for detecting microcystin (MC-LR) based on G-quadruplex nuclease, belonging to the technical field of immunoanalysis chemistry. Background technique [0002] The G-quadruplex is a nucleic acid sequence rich in guanine, a stable structure of four highly ordered nucleic acid strands formed by Hoogsteen-type base pairing. These quadruplex structures have been used as gene promoters because of their profound biological activities. In recent years, G-quadruplex-based DNases, complexes formed by hemin and G-quadruplexes, have shown surprising biocatalytic capabilities. Previous studies have found that G-quadruplex-based DNA enzymes have properties similar to peroxidases, which can effectively catalyze the oxidation of ABTS or TMB involving hydrogen peroxide, and finally form colored products. Therefore, G-quadruplex-based DNA enzymes are widely used in the fields of biomedicine and biocatalysis, such as the detection o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
Inventor 王立梅朱颖越朱益波齐斌
Owner CHANGSHU INSTITUTE OF TECHNOLOGY
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