Electrogenerated chemiluminescence bacterium sensing method and multi-functional probe

A multi-functional probe and electrochemistry technology, applied in the direction of chemiluminescence/bioluminescence, and analysis by making materials undergo chemical reactions, can solve the problems of expensive, complicated and cumbersome instruments and medicines, and achieve the goal of assembly process. Simple, large catalytic effect, good sensitivity

Active Publication Date: 2013-02-27
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

With the advancement of science and technology, a variety of new technologies have been used in the detection of bacteria, from early microscope observation and counting, enzyme-linked immunoassay, to the current differential pulse voltammetry, mass spectrometry, polymerase chain reaction, surface Enhanced Raman scattering, impedance, quartz crystal microbalance, surface plasmon resonance, etc., each method has shown unique advantages, but there are still shortcomings, such as most technical operations are cumbersome and complicated, and the instruments and drugs used are relatively expensive. The measurement process takes a long time, etc.

Method used

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  • Electrogenerated chemiluminescence bacterium sensing method and multi-functional probe
  • Electrogenerated chemiluminescence bacterium sensing method and multi-functional probe
  • Electrogenerated chemiluminescence bacterium sensing method and multi-functional probe

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Construction of an electrochemiluminescent method for the detection of Escherichia coli ATCC11775 based on a multifunctional probe:

[0030] a. Probe preparation:

[0031] All glassware used for the preparation of gold nanoparticles was first soaked in aqua regia for 24 h and then rinsed with deionized water. Add 92.5 milliliters of deionized water and 2.5 milliliters of chloroauric acid with a concentration of 4 grams per milliliter to a 250 milliliter three-neck flask, stir and heat until liquid droplets evenly return to the flask in the condenser tube. Add 5 ml of trisodium citrate dihydrate with a concentration of 10 mg per ml, stop heating after 15 minutes, then stir and cool to room temperature, and store at 4°C.

[0032] Before each experiment, take 3 ml with a concentration of 1.87×10 -9 Add 3 mg of glucose oxidase per liter of gold nanometer solution, and stir at room temperature for 24 hours. Centrifuge at 10,000 rpm for 10 minutes, wash with deionized wate...

Embodiment 2

[0041] Construction of an electrochemiluminescent method for detecting Pseudomonas aeruginosa ATCC27853 based on a multifunctional probe:

[0042] a. Probe Preparation

[0043] All glassware used for the preparation of gold nanoparticles was first soaked in aqua regia for 24 h and then rinsed with deionized water. Add 92.5 milliliters of deionized water and 2.5 milliliters of chloroauric acid with a concentration of 4 grams per milliliter to a 250 milliliter three-neck flask, stir and heat until liquid droplets evenly return to the flask in the condenser tube. Add 5 ml of trisodium citrate dihydrate with a concentration of 10 mg per ml, stop heating after 15 minutes, then stir and cool to room temperature, and store at 4°C.

[0044] Before each experiment, take 3 ml with a concentration of 1.87×10 -9 Add 3 mg of glucose oxidase per liter of gold nanometer solution, and stir at room temperature for 24 hours. Centrifuge at 10,000 rpm for 10 minutes, wash with deionized water,...

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Abstract

The invention belongs to the technical field of electrogenerated chemiluminescence and particularly relates to an electrogenerated chemiluminescence bacterium sensing method and a multi-functional probe. Modified glucose oxidase is gathered on nanogold particles, the multifunctional probe is built, and bacteria are detected through an electrogenerated chemiluminescence method. The glucose oxidase on the probe has double functions of identification and catalysis. First, the glucose oxidase is glycoprotein, carried oligosaccharide can be combined with lectin specificity, the glucose oxidase can catalyze glucose to oxidize to generate hydrogen peroxide, and the hydrogen peroxide can promote electrogenerated chemiluminescence of luminol. In addition, nanogold particles with large specific surface area have an effect of enriching enzyme and can catalyze luminol to emit light. The method is fast, simple, reliable, cheap and high in sensitivity and is used for detecting bacteria.

Description

technical field [0001] The invention belongs to the technical field of electrochemiluminescence, in particular to an electrochemiluminescence bacterial sensing method and a multifunctional probe. Background technique [0002] Bacteria are widely distributed in nature, are important participants in the earth's material cycle, and are closely related to human life. However, the existence of some pathogenic bacteria seriously affects food safety and water safety, causing various diseases and endangering human health. For example, the food poisoning outbreak in Europe in 2011 was caused by enteropathogenic Escherichia coli, which can cause severe diarrhea, sepsis, and even death. In addition, the widespread presence of Pseudomonas aeruginosa can infect wounds in many patients in hospitals, causing abscesses, lung infections, bacteremia, and sepsis, which are extremely harmful. Therefore, the detection, identification and quantification of bacteria are of great significance for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
Inventor 汪阳忠刘洋李景虹
Owner TSINGHUA UNIV
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