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Electrochemical method for detecting bacterial activity and concentration

A bacterial activity and electrochemical technology, applied in the field of electrochemical sensing, can solve the problems of time-consuming and high detection cost, and achieve the effect of low cost, obvious technical advantages, and detection limit

Active Publication Date: 2019-01-18
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Aiming at the time-consuming and high cost of detection existing in the detection of bacterial activity and concentration by fluorescent staining, enzyme-linked immunoassay, etc., the purpose of the present invention is to provide a simple, fast, high sensitivity, low detection limit, detection Wide range of methods available for electrochemical detection of bacterial activity and concentration

Method used

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  • Electrochemical method for detecting bacterial activity and concentration
  • Electrochemical method for detecting bacterial activity and concentration
  • Electrochemical method for detecting bacterial activity and concentration

Examples

Experimental program
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Effect test

Embodiment 1

[0036] An embodiment of the present invention for detecting bacterial activity, the schematic diagram of the principle is as follows figure 1 Shown. The detection method specifically includes the following steps:

[0037] (1) Mix the live bacteria solution and the dead bacteria solution in different proportions to obtain a series of standard E. coli solutions with different activities (0, 20%, 40%, 60%, 80% and 100%). After the bacterial solution was centrifuged, the supernatant was dropped onto the electrode surface;

[0038] (2) The electrode reacts with sodium molybdate: 1 mM sodium molybdate solution is added dropwise to the electrode surface to react at 25°C for 20 minutes.

[0039] (3) Square wave voltammetry detection: 0.5mol / L sulfuric acid solution is used as electrolyte, and the voltage range of 0-0.5V is measured at a frequency of 15Hz to obtain the peak current value of each electrode, and then each electrode The peak current value of and the corresponding bacterial act...

Embodiment 2

[0043] Staphylococcus aureus was used to replace Escherichia coli in Example 1, and the other operation steps were the same. The square wave voltammetry curve of the electrode detecting Staphylococcus aureus with different activity ratios is as figure 2 Shown.

Embodiment 3

[0045] An embodiment of the present invention for detecting the concentration of bacteria, its principle schematic diagram is as follows figure 1 Shown. The detection method specifically includes the following steps:

[0046] (1) After centrifuging the dead E. coli bacteria solution with different concentrations, a series of different concentrations (bacteria OD 600 The value of 0, 0.2, 0.35, 0.45, 0.65 and 0.75) standard E. coli solution, the supernatant was dripped onto the electrode surface;

[0047] (2) The electrode reacts with sodium molybdate: 1 mM sodium molybdate solution is added dropwise to the electrode surface to react at 25°C for 20 minutes.

[0048] (3) Square wave voltammetry detection: 0.5mol / L sulfuric acid solution is used as electrolyte, and the voltage range of 0-0.5V is measured at a frequency of 15Hz to obtain the peak current value of each electrode, and then each electrode Make a standard curve with the corresponding bacterial concentration of the peak curre...

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Abstract

The invention discloses an electrochemical method for detecting the bacterial activity and concentration. On the basis of the principle that bacteria can release phosphate-containing substances (suchas DNA and ATP) after being dead, the phosphate-containing substances can react with sodium molybdate to generate a current, and the current size is in inverse proportion to the bacterial activity andis in direct proportion to the bacterial concentration, the electrochemical detection method for the bacterial activity and concentration is constructed. The method has the advantages of being high in sensitivity, easy and rapid to operate, low in detection limit, wide in detection range and the like when being used for bacterial activity and concentration detection.

Description

Technical field [0001] The invention relates to a method for detecting the activity or concentration of bacteria, in particular to a method that uses the death of bacteria to release phosphate-containing substances (DNA, ATP, etc.), and the phosphate-containing substances can react with sodium molybdate to generate electric current, thereby A method for realizing electrochemical detection of bacterial activity and concentration; belongs to the field of electrochemical sensing technology. Background technique [0002] Bacteria have a great influence on human activities. On the one hand, bacteria are the pathogens of many diseases, including tuberculosis, gonorrhea, anthracnose, syphilis, plague, trachoma and other diseases are all caused by bacteria. However, humans often use bacteria. For example, the production of cheese, yogurt and fermented rice, the production of some antibiotics, and the treatment of waste water are all related to bacteria. In the field of biotechnology, b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48
CPCG01N27/48
Inventor 阳明辉蒋星星
Owner CENT SOUTH UNIV