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Electrochemical detection method for detecting kanamycin residues based on nucleic acid aptamer and nano analogue enzyme

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

Active Publication Date: 2016-07-20
上海谱尼医学检验实验室有限公司
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  • Abstract
  • Description
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  • 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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  • Electrochemical detection method for detecting kanamycin residues based on nucleic acid aptamer and nano analogue enzyme
  • Electrochemical detection method for detecting kanamycin residues based on nucleic acid aptamer and nano analogue enzyme
  • Electrochemical detection method for detecting kanamycin residues based on nucleic acid aptamer and nano analogue enzyme

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1. The drafting of kanamycin standard solution peak current value-concentration standard curve

[0032] Specific steps are as follows:

[0033] (1) Preparation of gold nanoparticle-mimicking enzyme: using tyrosine as reducing agent and trapping agent to prepare gold nanoparticle-mimicking enzyme, boil 300mL ultrapure aqueous solution containing 0.1mM tyrosine and 0.1mM KOH for 3-5min, immediately Add 1.02mL of chloroauric acid solution with a mass volume ratio of 1% and keep it in a boiling water bath for 5min; continue boiling in the boiling water bath until the solution volume is reduced to 30mL to obtain a gold nanoparticle solution; put the above gold nanoparticle solution in advance The excess KOH, metal ions and tyrosine were dialyzed in the dialysis bag with a molecular weight cut-off of 12kDa treated in boiling water to obtain the gold nanoparticle mimic enzyme, which was stored in a 4°C refrigerator for later use;

[0034](2) Preparation of function...

Embodiment 2

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

[0046] Here, a method of artificially polluting honey is prepared by adding standard concentration of kanamycin to honey to obtain a honey sample containing 0-600nM kanamycin residue.

[0047] The artificially contaminated kanamycin in acacia honey was detected according to the steps of standard substance detection in Example 1.

[0048] get as Figure 5 The shown differential pulse voltammetry detects the honey sample DPV current-potential curve, and Figure 4 The relationship curve between the peak current value and the concentration of kanamycin in the honey sample is shown.

[0049] In the range of kanamycin concentration of 1-60nM, there is a linear relationship between the peak current value and the concentration, and the linear regression equations are: 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 be...

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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 residue based on nucleic acid aptamers and nano-mimetic enzymes, belonging to the technical field of analytical chemistry. Background technique [0002] 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, Tuberculosis and some Gram-positive bacteria. It is widely used because of its low price, convenient administration and wide antibacterial spectrum It is used as a shared medicine for humans and animals with excellent effect. In agriculture, animal husbandry, and aquaculture, kanamycin can not only treat various infectious...

Claims

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Application Information

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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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