A biosensor for detecting Escherichia coli and its preparation method

A biosensor, Escherichia coli technology, applied in the field of sensors, can solve problems such as complicated instrument operation, achieve good repeatability, low process cost, and facilitate on-site detection

Active Publication Date: 2018-01-30
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the defect that the instrument operation is complicated and professional operators are required in the existing detection method, the present invention provides an electrochemical sensor for detecting Escherichia coli

Method used

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  • A biosensor for detecting Escherichia coli and its preparation method
  • A biosensor for detecting Escherichia coli and its preparation method
  • A biosensor for detecting Escherichia coli and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0054] A comparative experiment of a method for preparing an electrochemical sensor for detecting Escherichia coli, the steps are as follows:

[0055] a. Two gold electrodes are first polished in 0.3 and 0.05 μm alumina slurry until they become mirror surfaces, and then rinsed with secondary water;

[0056] b. Prepare 2% glutaraldehyde solution, use this solution to cross-link the E. coli antibody to the surface of the electrode, dry it for 60 minutes, and rinse the electrode with secondary water and PBS to remove unbound antibody;

[0057] c. After washing several times with secondary water, modify the bovine serum albumin behind the glutaraldehyde-E. coli antibody layer, so that the bovine serum albumin can block the non-specific binding site;

[0058] The detection method is as follows:

[0059] d. After fully agitating and washing in PBS buffer, incubate in E. coli bacteria solution, wash with PBS buffer, E. coli will bind to the electrode due to the specific binding betw...

Embodiment 3

[0067] A kind of stability analysis of the electrochemical sensor that detects escherichia coli, the steps are as follows:

[0068] a. The 4 gold electrodes are first polished in 0.3 and 0.05 μm alumina slurry until they become a mirror surface, and rinsed with secondary water;

[0069] b. Prepare 2% glutaraldehyde solution, use this solution to cross-link the E. coli antibody to the surface of the electrode, dry it for 60 minutes, and rinse the electrode with secondary water and PBS to remove unbound antibody;

[0070] c. After washing several times with secondary water, modify the bovine serum albumin behind the glutaraldehyde-E. coli antibody layer, so that the bovine serum albumin can block the non-specific binding site;

[0071] The detection method is as follows:

[0072] d. After the electrode is fully agitated and cleaned in PBS buffer, it is incubated in E. coli bacteria solution and washed with PBS buffer. E. coli is bound to the electrode due to the specific bindin...

Embodiment 4

[0079] A kind of standard curve drawing and practical application of the preparation method of the electrochemical sensor that detects Escherichia coli, the steps are as follows:

[0080] a. The 6 gold electrodes are first polished in 0.3 and 0.05 μm alumina slurry until they become a mirror surface, and rinsed with secondary water;

[0081] b. Prepare 2% glutaraldehyde solution, use this solution to cross-link the E. coli antibody to the surface of the electrode, dry it for 60 minutes, and rinse the electrode with secondary water and PBS to remove unbound antibody;

[0082] c. After washing several times with secondary water, modify the bovine serum albumin behind the glutaraldehyde-E. coli antibody layer, so that the bovine serum albumin can block the non-specific binding site;

[0083] The detection method is as follows:

[0084] d. After the electrode is fully agitated and washed in PBS buffer, it is 0, 1.8×10, 1.8×10 in E. coli bacteria solution 2 , 1.8×10 3 , 1.8×10 ...

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Abstract

The invention discloses a biosensor for detecting Escherichia coli, which consists of a gold electrode, a glutaraldehyde-Escherichia coli antibody layer, a bovine serum albumin sealing layer, an Escherichia coli layer and an RCA product layer from the inside to the outside. In the preparation method of the sensor, the biosensor electrode provided by the invention has stable performance, good repeatability, and a detection limit of 1.6×10 cfu mL‑1.

Description

technical field [0001] The invention relates to the technical field of sensors, in particular to an electrochemical sensor for detecting Escherichia coli, and also to a preparation method for the electrochemical sensor for detecting Escherichia coli. Background technique [0002] In recent years, the harm of pathogenic Escherichia coli to production and development has become more and more serious. The main reasons for the analysis are as follows: the primary disease caused by pathogenic E. coli is less, and the concurrent and secondary forms increase, which indirectly increases the harm of colibacillosis. Coliform bacteria are widely distributed and widely exist in feces of warm-blooded animals and in nature. Investigations and studies have shown that coliform bacteria mostly exist in the feces of warm-blooded animals, places where humans are frequently active, and places with feces pollution. The pollution of human and animal feces to the external environment is the main ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 王玉郭玉娜黄加栋刘素徐伟王虹智许颖邱婷婷崔洁
Owner UNIV OF JINAN
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