Electrochemical transducer, and preparation method and application for detecting pathogenic bacteria

A pathogenic bacteria and sensor technology, applied in the field of electrochemical sensors for detecting pathogenic bacteria, can solve problems such as time-consuming, time-consuming environmental requirements, and tediousness, and achieve the effects of improving detection sensitivity, easy popularization and application, and simple popularization and application

Active Publication Date: 2015-01-21
ANIMAL AND PLANT & FOOD DETECTION CENTER JIANGSU ENTRY EXIT INSPECTION AND QUARANTINE BUREAU
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, these methods are complicated to operate, have high requirements on the detection environment or instruments, and take a long time. For example, the plate method is time-consuming and has strict environmental requirements, and the instrument method is expensive and cumbersome.

Method used

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  • Electrochemical transducer, and preparation method and application for detecting pathogenic bacteria
  • Electrochemical transducer, and preparation method and application for detecting pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 detects the electrochemical sensor of Campylobacter jejuni

[0035] (1) Preparation of working solution 1

[0036] Working solution 1 is nano-magnetic bead dispersion labeled with anti-Campylobacter jejuni monoclonal antibody, the concentration of nano-magnetic beads is 10mmol / L, and the molar ratio of nano-magnetic beads to anti-Campylobacter jejuni monoclonal antibody is 1:6.

[0037] Dissolve 1.08 g of ferric chloride and 0.2 g of trisodium citrate in 20 mL of ethylene glycol, then add 1.2 g of sodium acetate, stir for 30 min, and place it in an oil bath at 200°C for 10 hours to obtain a concentration of 10 mM nano magnetic bead dispersion.

[0038] Take 60 μL of 10 mM nanomagnetic bead solution, 15 μL of 0.1 mM 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) solution and 0.01 mM N-hydroxy 15 μL of succinimide (NHS) solution, mixed, and reacted at room temperature for 10-30 minutes to activate the nano-magnetic beads; add 0.01 mg anti...

Embodiment 2

[0053] Example 2 Detection of Campylobacter jejuni in actual samples

[0054] 1. Establish a standard curve

[0055] The electrochemical sensor for detecting Campylobacter jejuni in Example 1 was used to detect Campylobacter jejuni. The detection process is at room temperature.

[0056] (1) Take 20 μL of working solution 1 and add it to 1 mL of milk solution supplemented with different concentrations of Campylobacter jejuni (ATCC33560, purchased from Beijing Zhongyuan Company). After mixing, place it on a rotary mixer. 10 minutes. The nanomagnetic beads-Campylobacter jejuni complex was magnetically separated by a magnetic stand, the supernatant was removed, 1 mL of 10 mM PBS (pH 7.4) solution was added, and washed on a rotary mixer for 3 minutes, and then repeated washing 3 times. Finally, 100 μL of PBS buffer (10 mM, pH 7.4) was added to resuspend to obtain the nanomagnetic beads-Campylobacter jejuni complex dispersion.

[0057] (2) Mix 20 μL immunomagnetic beads-Campylob...

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Abstract

The invention provides an electrochemical transducer, and a preparation method and application for detecting pathogenic bacteria, and relates to the field of pathogenic bacteria detection. The electrochemical transducer comprises a working electrode, wherein the working electrode is a wave carbon electrode with the surface modified with oligonucleotide probe 2. The electrochemical transducer further comprises a working solution 1 and a working solution 2, wherein the working solution 1 is a nanometer magnetic bead dispersion liquid marked with a pathogenic bacteria antibody; the working solution 2 is a gold nanometer particle dispersion liquid marked with a pathogenic bacteria antibody and oligonucleotide probe 1; the end 5 of the oligonucleotide probe 1 is modified with methylene blue, the end 3 of the oligonucleotide probe 2 is modified with amino, and the base sequences of the oligonucleotide probe 1 and the oligonucleotide probe 2 are complementary. According to the invention, the electrochemical transducer is convenient and rapid, and the detection sensitivity is significantly improved.

Description

technical field [0001] The invention relates to the field of pathogenic bacteria detection, in particular to an electrochemical sensor for detecting pathogenic bacteria, a preparation method and an application thereof. Background technique [0002] Pathogenic bacteria (Pathogenic bacteria) refers to microorganisms that can cause disease. Pathogenic bacteria include bacteria, viruses, spirochetes, rickettsia, chlamydia, mycoplasma, fungi, and actinomycetes. Generally speaking, pathogenic bacteria refer to bacteria in pathogenic microorganisms. The pathogenicity of bacteria is related to its virulence, invasion quantity and invasion portal. While the vast majority of bacteria are harmless or even beneficial, a significant number can cause disease. Opportunistic pathogens only cause disease under certain conditions, such as when there is a wound that allows bacteria to enter the bloodstream, or when immunity is weakened. [0003] For example: Campylobacter jejuni is a norma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 薛峰滕军蒋原陈伟曾德新蒋鲁岩袁芳夏晓莉邵景东
Owner ANIMAL AND PLANT & FOOD DETECTION CENTER JIANGSU ENTRY EXIT INSPECTION AND QUARANTINE BUREAU
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