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Aptamer wrapped gold nanoparticle probe-based reagent for colorimetric detection of omethoate and application of reagent

A gold nanoparticle and nucleic acid aptamer technology, which is applied in biological testing, analysis through chemical reaction of materials, material inspection products, etc., can solve the problems of harsh storage and activity conditions of enzyme molecules and affect the application of enzymes, and achieve sensitivity High and good detection rate, simple and easy detection method

Active Publication Date: 2015-08-19
迈科若(苏州)医疗科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the storage and activity conditions of enzyme molecules are relatively harsh, which affects the application of enzymes.

Method used

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  • Aptamer wrapped gold nanoparticle probe-based reagent for colorimetric detection of omethoate and application of reagent
  • Aptamer wrapped gold nanoparticle probe-based reagent for colorimetric detection of omethoate and application of reagent
  • Aptamer wrapped gold nanoparticle probe-based reagent for colorimetric detection of omethoate and application of reagent

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1: the preparation of reagent and its result

[0025] The preparation steps of omethoate reagent for colorimetric detection based on nucleic acid aptamer-coated gold nanoparticle probe are as follows: centrifuge and concentrate 1000 mL of gold nanoparticle solution to 500 μL, and the final concentration is about 2 nmol / L; slowly add 10 μL to a concentration of 100 μmol / L nucleic acid aptamer solution, its final concentration is 2 μ mol / L; After 1 hour of shaking reaction with 350rpm rotating speed, centrifuge with 10000rpm rotating speed for 15 minutes, add 0.01mol / L PBS (PH=7) resuspend, make reagent. Carry out ultraviolet detection to the prepared reagent, investigate its particle size change, its result is as follows: figure 2 As shown, a is the absorbance curve before the adsorption of the nucleic acid aptamer, and b is the absorbance curve after the adsorption of the nucleic acid aptamer, indicating that the particle size of gold nanoparticles does not...

Embodiment 2

[0026] Embodiment 2: the detection effect of kit colorimetric detection omethoate

[0027] The preparation and method of use of the kit: add salt solution to the mixture of omethoate and reagent, gold nanoparticle solution and reagent respectively, and then detect. Directly add 10 μl of NaCl solution with a concentration of 0.2M, let it stand for 1 minute, and perform UV-visible spectral absorption spectrum detection; directly add 10 μl of NaCl solution with a concentration of 0.2M to 90 μl of the reagent, and perform UV-visible spectral absorption spectroscopy after standing for 1 minute Detection: First add 10 μl of omethoate solution with a concentration of 50 μM to 90 μl of the reagent to obtain a mixture, then add 10 μl of a 0.2 M NaCl solution after standing for 10 seconds, and perform ultraviolet-visible absorption spectrum detection after standing for 1 minute. The test results are attached image 3 As shown, the absorption peak of the gold nanoparticle solution shown...

Embodiment 3

[0028] Example 3: Effect of random nucleic acid and nucleic acid aptamer on the detection effect of omethoate

[0029] The preparation and use method of the kit: prepare the control reagent with random nucleic acid of the same length as the nucleic acid aptamer, and prepare the reagent with the nucleic acid aptamer. The reagent preparation steps are as described in Example 1, wherein the control reagent is prepared with random nucleic acid instead of the nucleic acid aptamer , add 10 μl of omethoate solution with a concentration of 0 and 5 μM to 90 μl of the control reagent and the reagent respectively, mix well and let it stand for reaction for 10 seconds, then add 10 μl of a NaCl solution with a concentration of 0.2M, and carry out ultraviolet-visible analysis after standing for 1 minute. Spectroscopic absorption spectrum detection; the results are as follows Figure 4 As shown, the absorbance curve of the control reagent did not change significantly after adding 0 (a) and 5...

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Abstract

The invention discloses an aptamer wrapped gold nanoparticle probe-based reagent for the colorimetric detection of omethoate and application of the reagent. The reagent is prepared by the following steps of adding an aptamer solution at concentration of 2 to 5 mu M into a gold nanoparticle solution at concentration of 1 to 3nM, performing reaction for more than 1h, performing centrifugation at 8,000 to 12,000rpm, and performing resuspension to obtain the reagent by using a PBS solution at concentration of no higher than 0.1M, wherein a sequence of an aptamer is 5minute-AGCTTGCTGCAGCGATTCTTGATCGCCACAGAGCT-3minute. When the reagent is adopted for detecting omethoate, the lower limit of detection is lower than 0.2 mu M. Compared with a conventional omethoate detection reagent, the reagent is higher in sensitivity.

Description

technical field [0001] The invention belongs to the field of colorimetric reagent research, and relates to a reagent for colorimetric detection of omethoate based on a nucleic acid aptamer-wrapped gold nanoparticle probe and its application. Background technique [0002] Omethoate is widely used in agriculture, but its toxicity and irrational use also make it a large number of residues in water, soil, air and food, causing environmental pollution. Omethoate can inhibit the activity of neurotransmitter cholinesterase in blood, saliva and tissues, thereby damaging the nervous system of humans and animals, and endangering the health of humans and animals. [0003] There are many detection methods for omethoate residues at present, including liquid chromatography, gas chromatography, gas-liquid combination, mass spectrometry, surface-enhanced Raman spectroscopy, surface acoustic wave, etc. Although these detection methods have high sensitivity and reliability, But it requires c...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N33/53
Inventor 万莹王鹏娟苏岩杨树林
Owner 迈科若(苏州)医疗科技有限公司
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