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Tobramycin detection test paper based on aptamer and platinum modified gold nanoparticles

A technology of gold nanoparticles and tobramycin, which is applied in the fields of analytical chemistry, medicine, environment and food testing, can solve problems such as the limitation of visual detection of tobramycin, achieve quantitative analysis, visualize results, and improve detection efficiency. Effect

Active Publication Date: 2021-08-20
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection limit of tobramycin was 6.5×10 -9 mol / L, but the use of fluorescence limits the visual detection of tobramycin

Method used

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  • Tobramycin detection test paper based on aptamer and platinum modified gold nanoparticles
  • Tobramycin detection test paper based on aptamer and platinum modified gold nanoparticles
  • Tobramycin detection test paper based on aptamer and platinum modified gold nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Preparation of tobramycin detection test paper based on nucleic acid aptamer and platinum-modified gold nanoparticles, including preparation and functionalization of platinum-modified gold nanoparticles, pretreatment of gold standard pad and sample pad, and pretreatment of nitrocellulose membrane , Assembly of test strips, testing of test strips. The specific steps are:

[0056] (1) Preparation and functionalization of platinum-modified gold nanoparticles:

[0057] Soak the glassware used in aqua regia for 30 minutes, then thoroughly clean it with distilled water, then soak it in ultrapure water for 12 hours, and dry it for later use;

[0058] In the first step, the seeds of 15nm AuNPs were synthesized. Add 50 mL of chloroauric acid with a mass concentration of 0.01% into a 100 mL round bottom flask, stir and heat to boiling. Subsequently, 2 mL of a 1% sodium citrate aqueous solution was quickly added to the boiling solution. Keep the solution boiling and stirring f...

Embodiment 2

[0070] Determination of Tobramycin Standard Solution with Test Strip

[0071] Use 0.5mL centrifuge tubes to dilute tobramycin to different concentrations, 100μL per tube. Insert one end of the sample pad of the test strip prepared by the above method into the tobramycin standard solution of different concentrations, and react at room temperature for 5 minutes.

[0072] from figure 2 It can be seen from the low-sensitivity test results that when the concentration of tobramycin is between 1 and 150nmol / L, the color of the T line becomes darker with the increase of the concentration of tobramycin. When the concentration is 60nmol / L, it can be clearly observed by naked eyes. The detection line band was detected, and the color of the detection line basically did not change after the concentration was 150nmol / L. Therefore, the naked eye detection limit of the test paper to tobramycin in low sensitivity mode was 60nmol / L. The color development of T and C lines was scanned with a ...

Embodiment 3

[0075] Detection of Tobramycin Residues in Milk Samples

[0076] Here, artificially polluted milk was prepared by adding standard concentration tobramycin to milk to obtain 0.5nmol / L, 2.5nmol / L, 20nmol / L, 40nmol / L, 100nmol / L, 200nmol / L Tobramycin Vegetarian milk samples. For milk samples, the pH value was adjusted to 4.6 with 20% trichloroacetic acid, then incubated at 45°C for 10 min, and then centrifuged at 10,000 r / min for 20 min to remove coagulated protein and fat. Then follow the complete operation steps of the above-mentioned high-sensitivity detection mode to detect milk samples containing different concentrations of tobramycin. get Figure 4 Dipstick profiles of milk samples containing different concentrations of tobramycin are shown.

[0077] exist Figure 4 It can be seen that with the increase of tobramycin concentration, the color of the T line becomes darker, and when the tobramycin concentration reaches 40nmol / L, the color of the T line basically does not ch...

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Abstract

The invention relates to tobramycin detection test paper based on an aptamer and platinum modified gold nanoparticles. The test paper comprises a sample pad, a gold label pad, a nitrocellulose membrane, a water absorption pad, a PVC base plate, a detection line and a quality control line. Platinum-modified gold nanoparticles (Au@PtNPs) are used for loading a hybrid double strand of an aptamer (Apt) and a complementary probe (cDNA) to be used as a signal amplification label. The unique bifunctional properties (plasma optical property and nano mimic enzyme catalytic property) of Au@Pt NPs provide two different detection schemes: one is red only generated by the intrinsic color of AuNPs, and the other is more sensitive dark blue generated by a catalytic substrate, so that a detection mode adjusted as required is realized. The test paper strip provided by the invention is simple, convenient and rapid in use, is sensitive and efficient, and has huge application potential in field point detection of tobramycin residues in food and environment.

Description

technical field [0001] The invention relates to the fields of analytical chemistry, medicine, environment and food detection, in particular to a tobramycin detection test paper based on aptamers and platinum-modified gold nanoparticles. Background technique [0002] Tobramycin (Tobramycin) is a secondary metabolite produced by Streptomyces oberurus fermentation, molecular formula: C18H37N5O9, molecular weight 467.51, belongs to aminoglycoside antibiotics, has the characteristics of good water solubility, stable properties, broad antibacterial spectrum, etc., in clinical and It is widely used in animal medicine to treat infections caused by microorganisms such as Gram-negative bacteria. However, the excessive use of tobramycin in agriculture, animal husbandry, aquaculture and other fields will lead to its extensive residues in animal-derived foods and ecological environments, and eventually accumulate in the human body through the food chain. This may have a serious impact o...

Claims

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Application Information

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IPC IPC(8): C12Q1/6834C12Q1/6806G01N33/53
CPCC12Q1/6834C12Q1/6806G01N33/9446G01N33/5308C12Q2525/205C12Q2563/137C12Q2563/155C12Q2563/131C12Q2565/625
Inventor 周楠迪金鑫张雨婷王晓丽
Owner JIANGNAN UNIV
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