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A biosensor for detecting ampicillin and its preparation method and application

A biosensor, ampicillin technology, applied in the field of biosensors, can solve the problems of cumbersome sample pretreatment or concentration process, cumbersome operation, difficult to popularize, etc., and achieves the effects of short detection time, good repeatability and easy operation.

Active Publication Date: 2021-08-17
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the above problems that the method for detecting ampicillin in the prior art is cumbersome, time-consuming and laborious, the pretreatment or concentration process of the sample is cumbersome, and the instrument is expensive and difficult to popularize. A biosensor based on hyperbranched hybridization chain reaction of DNA Walker for ultra-sensitive detection of ampicillin with many kinds of substances, small molecular weight and simple synthesis process, also relates to its preparation method and application

Method used

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  • A biosensor for detecting ampicillin and its preparation method and application
  • A biosensor for detecting ampicillin and its preparation method and application
  • A biosensor for detecting ampicillin and its preparation method and application

Examples

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Effect test

Embodiment 1

[0064] The preparation method of described biosensor comprises the following steps:

[0065] (1) Preparation of gold nanoparticles;

[0066] (2) Modification of Walkker (containing -SH) and Track (containing -SH) to the surface of gold nanoparticles;

[0067] (3) Mixing the labeled nano-gold solution with the homogeneous reaction solution;

[0068] (4) Hybridization chain reaction, fluorescence detection.

[0069] The preparation method, preferably the preparation steps of gold nanoparticles are as follows:

[0070] (1) After installing the required instruments, add 200ml of ultrapure water into the three-necked flask (be careful not to let dust fall into the three-necked flask).

[0071] (2) Take 500μL (0.04g / ml) HAuCl 4 Put 500μl and 200ml ultrapure water into a single packaged centrifuge tube with a pipette gun, stir and heat at a stirring speed of about 450 rpm until boiling.

[0072] (3) Under the condition of stirring, take 3ml of 1% trisodium citrate solution and q...

Embodiment 2

[0090] A preparation method of the fluorescent biosensor of the present invention:

[0091] The preparation method of described biosensor comprises the following steps:

[0092] (1) Preparation of gold nanoparticles;

[0093] (2) Modification of Walkker (containing -SH) and Track (containing -SH) to the surface of gold nanoparticles;

[0094] (3) Mix the labeled gold nanoparticles solution with the homogeneous reaction solution.

[0095] (4) Hybrid chain reaction, fluorescence detection

[0096] The preparation method, preferably the preparation steps of gold nanoparticles are as follows:

[0097] (1) After installing the required instruments, add 200ml of ultrapure water into the three-necked flask (be careful not to let dust fall into the three-necked flask).

[0098] (2) Take 500μL (0.04g / ml) HAuCl4 in a single-package centrifuge tube, use a pipette gun to take 500μl and 200ml ultrapure water, stir and heat at a stirring speed of about 450 rpm, until boiled.

[0099] (...

Embodiment 3

[0117] The preparation method of described biosensor comprises the following steps:

[0118] (1) Preparation of gold nanoparticles;

[0119] (2) Modification of Walkker (containing -SH) and Track (containing -SH) to the surface of gold nanoparticles;

[0120] (3) Mix the labeled gold nanoparticles solution with the homogeneous reaction solution.

[0121] (4) Hybrid chain reaction, fluorescence detection

[0122] The preparation method, preferably the preparation steps of gold nanoparticles are as follows:

[0123] (1) After installing the required instruments, add 200ml of ultrapure water into the three-necked flask (be careful not to let dust fall into the three-necked flask).

[0124] (2) Take 500μL (0.04g / ml) HAuCl4 in a single-package centrifuge tube, use a pipette gun to take 500μl and 200ml ultrapure water, stir and heat at a stirring speed of about 450 rpm, until boiled.

[0125] (3) Under stirring conditions, take 3ml of 1% trisodium citrate solution and quickly ad...

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Abstract

The invention relates to the technical field of biosensors, in particular to a fluorescent biosensor based on hybridization chain reaction amplification. In order to solve the problems of relatively low specificity and sensitivity and high cost of the method for detecting ampicillin in the above prior art. A biosensor based on nucleic acid aptamer to detect ampicillin, the nicking endonuclease Nb.BbcCI cooperates with the chain hybridization chain reaction to realize cyclic amplification, and the combination of thioflavin T and G-quadruplex produces fluorescence, both phase reaction mixture. Preparation method: prepare gold nanoparticles; modify Walker and Track on the surface of gold nanoparticles; mix the labeled nano-gold solution with a homogeneous reaction solution; hyperbranched hybridization chain reaction, fluorescence detection; use the specific type of nucleic acid aptamer Recognition, the use of nucleic acid aptamers to detect the target ampicillin with high specificity; the use of hyperbranched hybridization chain reaction amplification to achieve signal amplification.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a biosensor for detecting ampicillin, a preparation method and application thereof. Background technique [0002] Ampicillin (ampicillin, AMP) is a kind of β-lactam antibiotics. It is a semi-synthetic penicillin that introduces an amino group at the α-position of the carboxyl group of the penicillin-side chain to change its polarity. Ampicillin is acid-resistant and enzyme-resistant, which overcomes the disadvantage that natural penicillin is not suitable for oral administration. Therefore, it is often used as the first choice for anti-inflammation and anti-infection in the process of animal husbandry. It can inhibit Gram-negative bacteria and positive bacteria to varying degrees. [0003] At present, among the analysis methods of ampicillin, commonly used AMP detection methods include high-performance liquid chromatography (HPLC), gas chromatography (GC), liquid chromatograph...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 刘素瞿晓南黄加栋王玉张儒峰赵一菡李莎莎
Owner UNIV OF JINAN
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