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Aptamer colloidal gold lateral chromatography test paper for detecting kanamycin

A kanamycin and lateral chromatography technology, which is used in food safety, nano-biosensing, analytical chemistry, medicine, and the environment. problem, to achieve the effect of high repeatability, high sensitivity detection, and improved hybridization efficiency

Pending Publication Date: 2021-07-20
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The strip requires AgNPs-DNA 1 Amplifying components, the structure of the test strip is relatively complicated, which is not conducive to the stable production of the product and the repeatability of the application

Method used

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  • Aptamer colloidal gold lateral chromatography test paper for detecting kanamycin
  • Aptamer colloidal gold lateral chromatography test paper for detecting kanamycin
  • Aptamer colloidal gold lateral chromatography test paper for detecting kanamycin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: A rapid detection test strip for kanamycin based on nucleic acid aptamer

[0050] Preparation and functionalization of gold nanoparticles, construction of streptavidin-biotin-capApt and streptavidin-biotin-capDNA composite structures, assembly of nucleic acid aptamer test strips. The specific steps are:

[0051] 1. Preparation and functionalization of gold nanoparticles

[0052] (1) Preparation of gold nanoparticles (AuNPs)

[0053] The glassware used for the synthesis and storage of nanomaterials in the experiment was soaked in aqua regia (hydrochloric acid: nitric acid = 3:1) for 12 hours, washed with ultrapure water before use.

[0054] AuNPs were prepared by sodium citrate reduction method. details as follows:

[0055] 1) Dissolve 100mL of 0.01% HAuCl 4 Add it into a 250mL Erlenmeyer flask, heat and stir until the solution bumps, and keep for 1-2min.

[0056] 2) Quickly add 2 mL of 1% trisodium citrate solution to the Erlenmeyer flask, and continue ...

Embodiment 2

[0073] Embodiment two: with test strip to the mensuration of kanamycin standard solution

[0074] (1) Preparation of Kanamycin Standard Solution

[0075] Kanamycin standard solution was diluted with Running buffer (4×SSC, pH7) to a final concentration of 5, 8, 5, 25, 50, 125 and 250 ng / mL. The nucleic acid aptamer (Aptamer) was diluted to 1 μM with ultrapure water.

[0076] (2) Establishment of standard curve for detection of kanamycin nucleic acid test strips:

[0077] Mix 99 μL of kanamycin standard solution with different concentrations and 1 μL of Aptamer solution and incubate for 20 min. After mixing and reacting, add the mixed solution to the sample pad for detection. After 3 min of reaction, measure (T / C) relative signal intensity and establish (T / C) C) Standard curve of the corresponding relationship between relative light signal intensity and different kanamycin concentrations.

[0078] The result is as image 3 As shown, when the concentration of kanamycin is 15n...

Embodiment 3

[0080] Example three: detection of kanamycin residues in milk samples

[0081] To test the recovery rate using a milk mock sample, the steps are:

[0082] (1) Pretreatment of AuNPs@polyA-DNA solution: Add 0.8-1.4 μL of the prepared and stored AuNPs@polyA-DNA onto the gold standard pad of the test strip, and store at 4°C.

[0083](2) Sample pretreatment: the milk sample was diluted 10 times and filtered through a 0.22 μm microporous membrane. Add different concentrations of kanamycin (50, 150, 250ng / mL) to the milk;

[0084] (3) Determination of the recovery rate of kanamycin in milk: 1 μL Aptamer (1 μM) was mixed with 99 μL milk solution containing different concentrations of kanamycin and incubated for 20 min, and then tested with test strips after the mixed reaction.

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Abstract

The invention discloses aptamer colloidal gold lateral chromatography test paper for detecting kanamycin, and belongs to the fields of analytical chemistry, medicine, environment, food safety, nano biosensing and the like. According to the invention, the complementary sequences (Aptamer and CapApt) of DNA can be rapidly and sensitively captured by utilizing AuNPs@ polyA-DNA, and the colloidal gold lateral chromatography test strip which is rapid, sensitive and low in cost is developed. The kanamycin detection by the test strip method is simple, convenient and rapid, the detection can be carried out at any time and anywhere, the test result can be observed only by adding a test solution into a sample port and completely developing the test strip after 5 minutes, and the detection efficiency can be greatly improved. Qualitative analysis is carried out by naked eyes, and quantitative analysis is carried out by using a colloidal gold test paper quantitative analyzer. The aptamer colloidal gold lateral chromatography test paper has important significance on kanamycin residue detection in food.

Description

technical field [0001] The invention relates to an aptamer colloidal gold lateral flow chromatography test paper for detecting kanamycin, which belongs to the fields of analytical chemistry, medicine, environment, food safety, nanobiological sensing and the like. Background technique [0002] Antibiotics are secondary metabolites produced by bacteria, fungi, animals or plants, and have various antipathogenic activities. Among them, kanamycin is an aminoglycoside antibiotic, which is widely used because of its strong water solubility, broad-spectrum antibacterial effect on many Gram-negative bacteria and Gram-positive bacteria, cheap price and convenient administration. Used in veterinary medicine and feed additives. However, the overuse of antibiotics will cause great problems to human health and environmental safety. Generally speaking, the abuse of antibiotics will lead to the increase of bacterial resistance and other side effects, such as ototoxicity, nephrotoxicity an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52
CPCG01N33/526
Inventor 彭池方常芮李秀萍
Owner JIANGNAN UNIV
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