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Method for rapidly detecting bacteria by using electrochemical method

An electrochemical and bacterial technology, applied in the field of rapid detection of bacteria by electrochemical methods, can solve the problems of expensive instruments, complicated operation, poor specificity and selectivity, and achieve good detection activity, high specificity and sensitivity, and good detection The effect of discriminative ability

Inactive Publication Date: 2013-09-18
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the detection time of the above method is shortened, the immunological reagents required are expensive and usually highly toxic, and the instruments required are also expensive, and the operation is complicated, and the specificity and selectivity are poor.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1.1 Preparation of probe molecule solution: Prepare several probe molecule derivatives into high-concentration mother solutions with PBS, and then dilute them with PBS to a concentration of 1*10 -4 mol / l, 4*10 -4 mol / l, 8*10 -4 mol / l, 12*10 -4 mol / l,. Drug probes were stored at -4°C. Before the experiment, the probe drug was taken out and shaken, and the electrochemical experiment was carried out.

[0022] 1.2 Experimental strains and culture methods: Standard strains: Escherichia coli (ATCC8739), Staphylococcus aureus (CGMCC1.89), Klebsiella pneumoniae (ATCC700600), Acinetobacter baumannii (ATCC19606), Proteus mirabilis (ATCC12453 )Wait. Clinical drug-resistant strains: Staphylococcus aureus (SA321), Klebsiella pneumoniae (KP450), Acinetobacter baumannii (AB135), Proteus mirabilis (PM102). LB liquid medium: 1% (w / v) tryptone, 0.5% (w / v) yeast extract, 1% (w / v) sodium chloride. NaOH adjusted the pH to 7.2-7.4. The LB agar plate is obtained by adding 0.5% (w / v) a...

Embodiment 2

[0026] 1.1 Preparation of probe molecule solution: Prepare several probe molecule derivatives into high-concentration mother solutions with PBS, and then dilute them with PBS to a concentration of 1*10 -4 mol / l, 4*10 -4 mol / l, 8*10 -4 mol / l, 12*10 -4 mol / l,. Drug probes were stored at -4°C. Before the experiment, the probe drug was taken out and shaken, and the electrochemical experiment was carried out.

[0027]1.2 Experimental strains and culture methods: Standard strains: Escherichia coli (ATCC8739), Staphylococcus aureus (CGMCC1.89), Klebsiella pneumoniae (ATCC700600), Acinetobacter baumannii (ATCC19606), Proteus mirabilis (ATCC12453 )Wait. Clinical drug-resistant strains: Staphylococcus aureus (SA321), Klebsiella pneumoniae (KP450), Acinetobacter baumannii (AB135), Proteus mirabilis (PM102). LB liquid medium: 1% (w / v) tryptone, 0.5% (w / v) yeast extract, 1% (w / v) sodium chloride. NaOH adjusted the pH to 7.2-7.4. The LB agar plate is obtained by adding 0.5% (w / v) ag...

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Abstract

The invention relates to a method for rapidly detecting bacteria by using an electrochemical method. The method comprises the following steps: dropwise adding a bacterial solution on an electrode by applying supramolecular recognition and using a carborane derivative or adriamycin having a stable electrochemical activity as a bacterial marker probe molecule, then immediately dropwise adding the marker probe molecule so as to carry out specific binding with bacteria, and detecting a differential pulse response signal; or based on a bacterial suspension as electrolyte, adding the marker probe molecule into the bacterial suspension by applying supramolecular recognition and using the carborane derivative or adriamycin having the stable electrochemical activity as the bacterial marker probe molecule, so as to carry out specific binding with bacteria, and then detecting an alternating current impedance curve. By using the method disclosed by the invention, the effects of counting bacteria, identifying species of bacteria and distinguishing drug-resistant strains and sensitive strains in a clinical sample are achieved, and the method is strong in detection universality, quick in speed, high in sensitivity, good in specificity, and high in detection efficiency; and the application of expensive immune reagents or detection assay kits is avoided.

Description

technical field [0001] The invention relates to a method for rapidly detecting bacteria by using an electrochemical method. Background technique [0002] Pathogenic bacteria have always been "invisible killers" that seriously endanger human health. In order to reduce its harm to humans, it is necessary to carry out scientific research such as counting of viable bacteria, classification and identification, sensitivity, etc. At present, the most commonly used and relatively accurate counting method for viable bacteria is the agar plate colony counting method. This method is to dilute the sample to an appropriate multiple and culture it for 24 hours, so that the bacteria can be cultured to form discrete colonies, and then count manually. The identification of bacterial species will be carried out through a series of biochemical tests after the above culture to achieve. This method not only takes a long time to detect, but also wastes a lot of manpower and material resources. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26
Inventor 王雪梅吕夏毅李水红周研研
Owner SOUTHEAST UNIV