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An electrochemical detection method for the detection of kanamycin residues based on nucleic acid aptamers and nano-mimetic enzymes

A nucleic acid aptamer, kanamycin technology, applied in the field of analytical chemistry, can solve the problems of large result error, complicated operation, expensive equipment, etc., and achieve the effects of easy preparation, high sensitivity and high specificity

Active Publication Date: 2018-09-21
上海谱尼医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The microbiological detection method is widely used, and its advantage is that it is low in cost and can be operated by general laboratories, but the measurement time is long, the result error is large, the operation is complicated, and it cannot meet the needs of specificity and quantitative detection.
The instrumental inspection method uses the structure and physical and chemical properties of kanamycin for separation and qualitative and quantitative determination. It has good accuracy, but the equipment is often expensive, and it also has high requirements for experimenters and high requirements for sample pretreatment. Analysis Time-consuming and difficult to promote

Method used

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  • An electrochemical detection method for the detection of kanamycin residues based on nucleic acid aptamers and nano-mimetic enzymes
  • An electrochemical detection method for the detection of kanamycin residues based on nucleic acid aptamers and nano-mimetic enzymes
  • An electrochemical detection method for the detection of kanamycin residues based on nucleic acid aptamers and nano-mimetic enzymes

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1. Drawing of kanamycin standard solution peak current value-concentration standard curve

[0032] Specific steps are as follows:

[0033] (1) Preparation of gold nanoparticle mimic enzyme: use tyrosine as reducing agent and capture agent to prepare gold nanoparticle mimic enzyme, boil 300mL ultrapure aqueous solution containing 0.1 mM tyrosine and 0.1 mM KOH for 3-5 minutes, Immediately add 1.02 mL of a 1% chloroauric acid solution by mass to volume and keep it in a boiling water bath for 5 minutes; continue boiling in the boiling water bath until the volume of the solution is reduced to 30 mL to obtain a gold nanoparticle solution; advance the above gold nanoparticle solution Dialysis in a dialysis bag with a molecular weight cut-off of 12kDa treated in boiling water to remove excess KOH, metal ions and tyrosine to obtain a gold nanoparticle mimic enzyme, which is stored in a refrigerator at 4°C for later use;

[0034] (2) Preparation of functionalized gold nanopart...

Embodiment 2

[0045] Example 2. Detection of Kanamycin Residues in Honey Samples

[0046] Here, a standard concentration of kanamycin was added to honey to prepare artificially polluted honey, and a honey sample containing 0-600 nM kanamycin residue was obtained.

[0047] According to the standard detection procedure in Example 1, the artificially contaminated kanamycin in acacia honey was detected.

[0048] Get as Figure 5 Shown is the DPV current-potential curve diagram of honey samples detected by differential pulse voltammetry, and Figure 4 The curve showing the relationship between the peak current value and the concentration of kanamycin in the honey sample.

[0049] In the kanamycin concentration range of 1-60 nM, there is a linear relationship between the peak current value and the concentration. The linear regression equations are respectively: y=1.41579+0.0182x, R 2 =0.9978, where y is the peak current value (μA), and x is the concentration of kanamycin in the honey sample (nM). It can...

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Abstract

The invention provides an electrochemical detection method for detecting kanamycin residues based on a nucleic acid aptamer and a nano analogue enzyme and belongs to the technical field of analytical chemistry. Gold nano-particles are synthesized by reducing chloroauric acid through tyrosine, and hydrogen peroxide and reduced-state thionine are catalyzed to react to generate oxidized-state thionine; and the thionine can be detected through a difference pulse voltammetry. The gold nano-particles are modified through utilizing a kanamycin specific aptamer; and the aptamer is adsorbed on the surfaces of the gold nano-particles so that the peroxidase activity is inhibited. When a target object kanamycin exists, the aptamer can be competitively replaced from the surfaces of the gold nano-particles to form a compound, so that the peroxidase activity is recovered. The detection of the kanamycin can be realized through detecting a relation between a reduction peak current value of the oxidized-state thionine and an antibiotic concentration by utilizing the difference pulse voltammetry. The method provided by the invention has good repeatability, good stability and high sensitivity, and can be used for effectively detecting kanamycin residues in food samples.

Description

Technical field [0001] The invention relates to an electrochemical detection method for detecting kanamycin residues based on nucleic acid aptamers and nano mimic enzymes, and belongs to the technical field of analytical chemistry. Background technique [0002] Kanamycin (kanamycin) is a broad-spectrum aminoglycoside antibiotic with molecular formula C 18 H 38 N 4 O 15 S, molecular weight 582.58. Kanamycin is mainly extracted from the fermentation broth of Streptomyces griseus or Micromonas or semi-synthetically extracted from natural products. As a water-soluble antibiotic, Kanamycin sulfate is effective against many Gram-negative bacteria including Salmonella, Mycobacterium tuberculosis and certain Gram-positive bacteria. Because of its low price, convenient medication, and wide antibacterial spectrum It is used as a common medicine for humans and animals with excellent effect. In agriculture, animal husbandry and aquaculture, kanamycin can not only treat various infectious d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26G01N27/30
CPCG01N27/26G01N27/30G01N27/301
Inventor 周楠迪王春帅田亚平
Owner 上海谱尼医学检验实验室有限公司
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