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Method for rapidly detecting antibiotics based on colorimetric aptamer sensor

An aptamer sensor and antibiotic technology, which is applied in color/spectral characteristic measurement, instruments, measuring devices, etc., can solve the problems of low detection limit, inability to realize rapid on-site detection, and insufficient detection range, so as to improve sensitivity and detect Accuracy and sensitivity improvement, high sensitivity effect

Pending Publication Date: 2021-03-09
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The current colorimetric aptasensors still have the problems of insufficient low detection limit, insufficient detection range and inability to achieve rapid on-site detection.

Method used

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  • Method for rapidly detecting antibiotics based on colorimetric aptamer sensor
  • Method for rapidly detecting antibiotics based on colorimetric aptamer sensor
  • Method for rapidly detecting antibiotics based on colorimetric aptamer sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The detection of embodiment 1 kanamycin

[0038] (1) Aldezylation of Amino Magnetic Beads

[0039] That is, amino magnetic beads are activated, and the specific steps are as follows: use a pipette gun to pipette 3 mL (150 mg) of 8600-10 amino magnetic beads (particle size 1.5 μm) purchased from Polysciences, Inc. in the United States into a 50 mL centrifuge tube; add 24 mL of 0.01 M Pyridine buffer, shake vigorously at room temperature and mix evenly, carry out magnetic separation, discard the supernatant, this process is repeated three times; then add 12mL of 5% (volume fraction) glutaraldehyde solution (diluted with 0.01M pyridine buffer preparation), shake in a non-magnetic mixing device, i.e., a constant temperature shaker, at 25°C for 3 hours in the dark to ensure that the magnetic beads and glutaraldehyde solution are fully mixed; then perform magnetic separation to remove unreacted glutaraldehyde solution, and use 24mL Wash with 0.01M pyridine buffer repeatedly ...

Embodiment 2

[0063] Select the aptamer sequence that can specifically recognize streptomycin. The control group is to incubate the aptamer sequence, deionized water and the pre-prepared magnetic beads-streptomycin magnetic complex together, and the aptamer will interact with The magnetic beads-streptomycin fully reacted; in the experimental group, the aptamer sequence and different concentrations of free streptomycin were incubated with the pre-prepared magnetic beads-streptomycin magnetic complex, and the free streptomycin would interact with the Magnetic beads-streptomycin jointly competed for aptamer, so that the aptamers immobilized on the magnetic bead complex decreased. All the other operations are the same as in Example 1.

[0064] The aptamer (Streptomycin aptamer) specifically recognizing streptomycin was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and its sequence is as follows:

[0065] 5′-TAG GGA ATT CGT CGA CGG ATC CGG GGT CTG GTG TTC TGC TTT GTT CTGTCG GGT CGT...

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Abstract

The invention belongs to the field of food safety detection, and particularly relates to a method for rapidly detecting antibiotics based on a colorimetric aptamer sensor, which comprises the following steps of: (1) mixing and incubating a magnetic particle compound, an aptamer of target antibiotics and a sample to be detected or a standard solution, carrying out magnetic separation, and taking the magnetic particle compound combined with the aptamer, the magnetic nano-composite being a magnetic bead connected with a target antibiotic to be detected; (2) carrying out TdT amplification reactionon the magnetic particle compound combined with the aptamer by using deoxynucleotide and biotin modified deoxynucleotide, extending at the 3' -OH end of the aptamer, and carrying out magnetic separation to obtain a magnetic nano-compound subjected to TdT amplification; (3) connecting the product obtained in the step (2) with avidin-modified horseradish peroxidase to obtain a colorimetric aptamersensor; and (4) adding hydrogen peroxide and TMB (tetramethylbenzidine) into the colorimetric aptamer sensor, carrying out magnetic separation after reaction in a dark place, and taking the supernatant to detect the absorbance. The method can amplify the colorimetric detection signal so as to improve the detection sensitivity.

Description

technical field [0001] The invention relates to the field of food safety detection, and specifically discloses a colorimetric aptamer sensor based on TdT enzyme and magnetic beads and a method for rapidly detecting antibiotics by using the sensor. Background technique [0002] Antibiotics are a class of chemical substances that can affect normal life activities such as the growth and development of other cells. The currently commonly used antibiotics mainly include: chloramphenicols, aminoglycosides, macrolides, β-lactams, lincomycins, quinolones, tetracyclines, etc. As a class of anti-infection agents or antibacterial and growth-promoting feed additives, antibiotics have played an active role in food-related industries such as animal husbandry and fishery. However, with the frequent occurrence of antibiotic abuse, especially the detection of antibiotics in food and drinking water, people are paying more and more attention to this problem. Antibiotic residues in food is a ...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/543G01N21/31
CPCG01N33/5308G01N33/54326G01N33/54346G01N21/31
Inventor 颜娟赵婷婷张彤陶晴张倩雯
Owner SHANGHAI OCEAN UNIV
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