Method for detecting content of aminoglycoside antibiotics in solution to be detected

A technology of aminoglycosides and antibiotics, applied in measuring devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of continuous on-line detection of targets, no quantitative response, poor interface regeneration performance, etc., and achieve fast and low cost. , the effect of stable performance

Inactive Publication Date: 2018-06-12
TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, on the one hand, these nucleic acid aptamer sensors are limited by the poor interface regeneration performance, and cannot complete the continuous online detection of the target; For other aminoglycoside antibiotics (such as kanamycin B, gentamicin and amikacin), there is no quantitative response

Method used

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  • Method for detecting content of aminoglycoside antibiotics in solution to be detected
  • Method for detecting content of aminoglycoside antibiotics in solution to be detected
  • Method for detecting content of aminoglycoside antibiotics in solution to be detected

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Experimental program
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Effect test

Embodiment 1

[0063] Embodiment 1, the method for detecting the content of aminoglycoside antibiotics in the solution to be tested

[0064] The inventor of the present invention has found a method for detecting the content of aminoglycoside antibiotics in the solution to be tested through a large number of experiments. The specific principle diagram is shown in figure 1 : When aminoglycoside antibiotics do not exist, FAP is easy to form nucleic acid aptamer intermolecular multimer structure (ie M-Apts); when aminoglycoside antibiotics exist, aminoglycoside antibiotics can specifically bind to FAP and destroy polymer structure and form a single-chain hairpin structure with fluorescence quenching effect; graphene oxide (GO) can selectively adsorb and quench the single-chain hairpin structure in solution, which further enhances the effect on the single-chain hairpin structure The quenching efficiency is beneficial to increase the sensitivity of the all-fiber evanescent wave biosensor. Without...

Embodiment 2

[0143] Embodiment 2, selective detection

[0144] 1. Preparation of AAP probe modified optical fiber

[0145] Same as Step 1 in Step A of Example 1.

[0146] 2. Selective detection of different antibiotics

[0147] Set up the experimental group and conduct three repeated experiments, and the steps of each repeated experiment are as follows:

[0148] 1. Acquisition of nucleic acid aptamer multimer structure (M-Apts)

[0149] Same as 1 in Step A, Step 2 of Example 1.

[0150] 2. Combine M-Apts with samples

[0151] (1) Get the antibiotic to be tested (kanamycin A, amikacin, tetracycline, streptomycin, chloramphenicol or sulfamethazine), and then use pH7.4, 10mM Tris-HAc buffer dilution, A solution of the antibiotic to be tested with a concentration of 400 μM was obtained.

[0152] (2) Take the M-Apts formed in step 1 and dilute it with a pH 7.4, 10 mM Tris-HAc buffer to obtain an M-Apts solution with a concentration of 40 nM.

[0153] (3) Take a centrifuge tube, add 500 μ...

Embodiment 3

[0161] Embodiment 3, repeatability detection

[0162] 1. Preparation of AAP probe modified optical fiber

[0163] Same as Step 1 in Step A of Example 1.

[0164] 2. Repeatability detection

[0165] 1. Acquisition of nucleic acid aptamer multimer structure (M-Apts)

[0166] (1) Take pH 7.4, 10mM Tris-HAc buffer containing 1μM FAP, bathe in water at 95°C for 10min (for denaturation), then immediately transfer to ice bath at 0°C for 15min (for the purpose of combining FAP molecules with each other) ), get M-Apts.

[0167] (2) Take the M-Apts formed in step 1 and dilute it with a pH 7.4, 10 mM Tris-HAc buffer to obtain a 20 nM M-Apts solution.

[0168] 2. Signal detection

[0169] (1) Install the AAP probe-modified optical fiber prepared in step 1 into the all-fiber evanescent wave biosensor.

[0170] (2) Take the M-Apts solution and pass it into the all-fiber evanescent wave biosensor that completed the step (1), and detect the fluorescence signal at 500 s.

[0171] (3) Af...

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Abstract

The invention discloses a method for detecting the content of aminoglycoside antibiotics in a solution to be detected. The method disclosed by the invention adopts a kanamycin A aptamer and an AAP probe to specifically recognize other aminoglycoside antibiotics so as to realize detection of multiple substances on a single sensor. FurthermoreMeanwhile, the method disclosed by the invention has no need of an additional signal separation step, signal detection can be directly performed after reaction, and a sensor modified method ensures the stability and reproducibility of quantitative detection. The method disclosed by the invention is simple, obtains stable performance and good optical fiber repeatability are obtained, and the method is suitable for detection of aminoglycoside antibioticsin environmental samples as well as industrialization requirements of biosensors, and has important application values.

Description

technical field [0001] The invention belongs to the technical field of environmental monitoring and environmental ecological protection, and in particular relates to a method for detecting the content of aminoglycoside antibiotics in a solution to be tested. Background technique [0002] Antibiotics play an important role in the treatment of infectious diseases, but due to serious abuse in recent years, the problem of antibiotic residues in animal foods and environmental media (such as water and soil) has become prominent. These residual antibiotics will accumulate in the natural environment and in the human body, causing the human body to produce drug-resistant strains, or produce toxic effects on the body due to a large amount of accumulation. [0003] Antibiotics can often be classified according to their molecular structure. Aminoglycoside antibiotics are currently the most widely used antibiotics, such as kanamycin A, kanamycin B, gentamicin and amikacin. Abuse of ami...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/643
Inventor 何苗唐云飞韩世同
Owner TSINGHUA UNIV
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