Biosensor for detecting streptomycin based on aptamer and preparation method of biosensor

A biosensor and nucleic acid aptamer technology, applied in the field of biosensors, can solve the problems of difficult screening of bacteria, poor stability and low sensitivity of microbial analysis methods, and achieve the effects of fast detection speed, low detection limit and improved sensitivity

Active Publication Date: 2015-07-08
UNIV OF JINAN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, both methods have their own inevitable defects
Microbial analysis has poor stability, low sensitivity, and bacteria are not easy to screen; while HPLC requires more expensive sample pretreatment

Method used

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  • Biosensor for detecting streptomycin based on aptamer and preparation method of biosensor
  • Biosensor for detecting streptomycin based on aptamer and preparation method of biosensor

Examples

Experimental program
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Effect test

Embodiment 1

[0051] A preparation method of the electrochemical biosensor of the present invention:

[0052] a. The gold electrode is first polished in 0.3 and 0.05 μm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0053] b. Drop 10 μL of MB capture probe (10 μM) onto the electrode surface and incubate at room temperature for 2 hours. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0054] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0055] a. Mix sterilized water, 10X buffer (buffer), 1 μL ssDNA with a concentration of 50 μM, 1 μL hairpin-structured DNA chain (10 μM) and 1 μL different concentrations of target substances (100 pM, 500 pM, 1 nM, 5 nM, 10 nM, 50nM, 100nM, 500nM, 1μM) into pre-prepared sterilized centrifuge tubes. Shake for 30s, then put the mixed mix...

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Abstract

The invention provides a biosensor for detecting streptomycin based on an aptamer. The biosensor comprises a gold electrode, wherein an MB capture probe layer, HAP and an ssDNA layer are sequentially modified on the gold electrode. The invention also provides a preparation method of the biosensor. Based on specific recognition of the aptamer and the target, the sensor has the advantages of high detection speed, low detection limit, high specificity and the like, the defects of the conventional streptomycin detection method can be overcome, and rapid and accurate quantitative determination is realized.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a biosensor for detecting streptomycin based on nucleic acid aptamers, and also relates to a preparation method thereof. Background technique [0002] Streptomycin (SM), a common glucosamine-type antibiotic, is produced by soil actinomycetes and has significant antibacterial effects on a variety of Gram-positive and Gram-negative bacteria. Inhibits the growth and reproduction of bacteria (such as: tuberculosis, plague, whooping cough, bacillary dysentery, urinary tract infections and other infectious diseases mainly caused by Gram-negative bacteria). It can effectively prevent and control plant bacterial diseases, such as: agar fire blight, blueberry fungus, tobacco wildfire disease, etc., so it is often used in animal husbandry and aquaculture. However, the abuse of streptomycin in animal husbandry causes drug residues in agricultural products to cause some diseases. T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N27/327G01N33/543
Inventor 王玉崔洁黄加栋刘素徐伟王虹智郭玉娜许颖邱婷婷
Owner UNIV OF JINAN
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