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Preparation method and application of rapid detection kit for copper-containing nanorod compound

A detection kit and technology of copper nanorods are applied in the direction of material analysis, color/spectral property measurement, and chemical reaction of materials by observing the influence on chemical indicators, so as to achieve easy preparation and difficult inactivation. , the effect of good application prospects

Inactive Publication Date: 2019-01-18
HUNAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although clinical laboratories routinely measure insulin and glucose levels, until now researchers have rarely tested the two together

Method used

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  • Preparation method and application of rapid detection kit for copper-containing nanorod compound
  • Preparation method and application of rapid detection kit for copper-containing nanorod compound
  • Preparation method and application of rapid detection kit for copper-containing nanorod compound

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Experimental program
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Effect test

Embodiment 1

[0041] Firstly, copper-containing nanorod composites are prepared. Add 200 μL of 1M Na to 30 mL of the gold nanorod solution prepared by the seed crystal method 2 S 2 o 3 solution, placed in 50°C hot water and heated for 5 minutes to obtain core-shell gold nanorods, measure their corresponding absorption peaks on a UV-visible photometer, and the longitudinal plasma absorption peaks of gold nanorods red-shifted, continue to the solution Add 400μL 0.2M Cu(NO 3 ) 2 Solution, the longitudinal plasmon absorption peak of core-shell nanorods will blue-shift again to obtain copper-containing nanorod composites, such as figure 1 As shown in , after acting for 12 hours, centrifuge at 8000 rpm for 10 minutes on a centrifuge, remove the supernatant, and disperse with double distilled water for later use.

[0042] Prepare 14 centrifuge tubes, add 300 μL copper-containing nanorod complex to the centrifuge tubes, and then add 300 μL 5 mmol / L TMB and 300 μL 10 mmol / L H 2 o 2 , and then...

Embodiment 2

[0045] For the detection of glucose:

[0046] Add 200 μL of 1M Na to 30 mL of the gold nanorod solution prepared by the seed crystal method 2 S 2 o 3 solution, placed in hot water at 50°C for 5 minutes, and then added 400 μL of 0.2M Cu(NO 3 ) 2 Solution, after acting for 12 hours, centrifuge with 8000 rpm for 10 minutes on a centrifuge, remove the supernatant, and disperse it with twice distilled water for subsequent use. Add 300 μL of glucose solutions with concentrations of 0, 0.001, 0.01, 0.02, 0.4, 0.8, 2, 4, and 8 mM and 300 μL of 0.1 mg / mL glucose oxidase into 9 centrifuge tubes, and place them at 37°C for 30 minutes. Then add 300 μL copper-containing nanorod complex and 300 μL 5mmol / LTMB respectively, and finally adjust the pH to about 6 with 0.1mol / L HCl, and the color change is as follows: Figure 5 shown. After the color development is stable, measure its absorption spectrum on an ultraviolet spectrophotometer, and the ultraviolet absorption spectra of glucose ...

Embodiment 3

[0048] For the detection of insulin content:

[0049] Add 200 μL of 10 to 10 mL of copper-containing nanorod complex -4 μmol / L INS-aptamer, acted at 4°C for 12h, centrifuged at 4000rpm for 10min, re-dispersed with twice distilled water, and set aside.

[0050] Add 100 μL of the above-mentioned copper-containing nanorod complexes connected with INS-aptamer to each of the 8 centrifuge tubes, and then add 300 μL of the concentration of 0 and 10 -9 、10 -8 、10 -7 、10 -6 、10 -5 、10 -4 、10 -3 mg / mL insulin, after reacting for 20 minutes, add 200 μL of 10 mmol / L hydrogen peroxide solution for 30 minutes, and then add 300 μL of 5 mmol / L TMB respectively. After 30 minutes, its color changes as Figure 9 shown. The quantitative detection of insulin was performed in parallel three times, and the concentrations from left to right were: 0, 0.014, 0.054, 0.216, 0.270, 0.540, 2.16μU / mL, and the color changes as follows: Figure 10 shown. Under the conditions of different concentrat...

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Abstract

The invention discloses a preparation method and an application of a rapid detection kit for a copper-containing nanorod compound. The method comprises the following steps of preparing the copper-containing nanorod compound; adding glucose solutions with different concentrations into a centrifuge tube, adding glucose oxidase, adding the copper-containing nanorod compound and a color developing agent, adjusting the pH value, obtaining a standard curve with a good linear relation according to a relation between an absorbance value of the color developing agent at the wavelength of 652nm and theglucose concentration, and based on this, performing calculation to obtain the glucose content in a to-be-detected sample; and adding the copper-containing nanorod compound connected with an INS-aptamer into the centrifuge tube, adding insulin with different concentrations, adding a hydrogen peroxide solution and the color developing agent (TMB), obtaining a standard curve with a good linear relation according to a relation between the absorbance value of the color developing agent at the wavelength of 652nm and the insulin concentration, and based on this, performing calculation to obtain theinsulin content in the to-be-detected sample. The kit also can be used for distinguishing type 1 diabetes and type 2 diabetes. The method is obvious in phenomenon, easy and convenient in operation, high in sensitivity, and accurate and effective.

Description

technical field [0001] The invention belongs to the technical field of chemical biosensing and biological detection, and in particular relates to a preparation method and application of a copper-containing nanorod complex rapid detection kit with peroxide-mimicking enzyme properties. Background technique [0002] Diabetes is a global health problem that reduces the quality of life for many and is a major economic burden on society due to increased health costs and loss of economic activity. [0003] Diabetes mellitus is a chronic metabolic disease, which can be mainly divided into type 1 diabetes mellitus (T1DM) or type 2 diabetes mellitus (T2DM). T1DM has reduced or no insulin production due to the destruction of pancreatic β cells, while T2DM patients have low intracellular insulin sensitivity. Thus, both T1DM patients and T2DM patients are characterized by altered blood glucose levels. When insulin levels are low or absent, blood sugar concentrations increase, resulting ...

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 黄昊文谭芳汪志芳杨艳谢潇雪花欣怡阳秀梅
Owner HUNAN UNIV OF SCI & TECH
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