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A biosensor for detecting Salmonella typhimurium

A biosensor, Salmonella technology, applied in the field of biosensors, can solve the problems of complicated instrument operation and long time consumption, and achieve the effects of good repeatability, improved sensitivity, and accurate quantitative detection

Active Publication Date: 2019-03-26
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Aiming at the disadvantages of complex instrument operation, long time-consuming and professional operators in the existing detection methods, the present invention provides a cascading signal aided by exonuclease III with strong specificity, high sensitivity, low cost and fast detection speed Amplified electrochemical biosensor for the detection of Salmonella typhimurium

Method used

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  • A biosensor for detecting Salmonella typhimurium
  • A biosensor for detecting Salmonella typhimurium
  • A biosensor for detecting Salmonella typhimurium

Examples

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preparation example Construction

[0049] The preparation method of described biosensor comprises the following steps:

[0050] (1) Pretreatment of the electrodes;

[0051] (2) Modify the mixture of iDNA and Helper on the electrode surface;

[0052] (3) Modification of homogeneous reaction products onto the electrode surface.

[0053] In the preparation method, the operation steps of modifying the mixture of iDNA and Helper to the surface of the electrode are preferably as follows: add 10 μL of the mixture of iDNA and Helper dropwise to the pretreated electrode surface, and incubate at 37 °C for 2 h .

[0054] In the preparation method, the preferred steps of modifying the homogeneous reaction product onto the electrode surface are as follows:

[0055] (1) Add 14 μL of sterilized water, 2 μL of 10× buffer, 2 μL of 10 μM P1 and 2 μL of 10 μM P2 into a centrifuge tube, shake for 30 s, place in a thermostat at 90 °C for 10 min, and cool naturally to room temperature;

[0056] (2) Add 12 μL of sterilized water...

Embodiment 1

[0063] The main steps of the electrode modification process are as follows:

[0064] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0065] b. Drop 10 μL of the mixture of iDNA and Helper (10 μM) onto the electrode surface, and incubate at 37 °C for 2 h. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0066] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0067] a. Add sterilized water, 10× buffer, P1 and P2 (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 12 μM, 14 mM) into the centrifuge tube, shake for 30 s, put Annealed in an incubator at 90 °C for 10 min, then cooled naturally to room temperature;

[0068] b. Add sterilized water, 10× buffer, aptamer, P3 and the target object to be tested into the centrifuge ...

Embodiment 2

[0079] The main steps of the electrode modification process are as follows:

[0080] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0081] b. Drop 10 μL of the mixture of iDNA and Helper (10 μM) onto the electrode surface, and incubate at 37 °C for 2 h. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0082] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0083] a. Add sterilized water, 10× buffer, P1 and P2 into the centrifuge tube, shake for 30 s, put it in a thermostat at 90 ℃ for 10 min, and cool down to room temperature naturally;

[0084] b. Add sterilized water, 10× buffer, aptamer, P3 (2 nM, 4 nM, 6 nM, 8 nM, 10 nM, 12 nM, 14 nM) and the target substance to be tested into the centrifuge tub...

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Abstract

The invention provides a salmonella typhimurium detection biosensor. The salmonella typhimurium detection biosensor is prepared by sequentially modifying an electrode with an iDNA layer, a Helper layer and homogeneous reaction mixed liquor. The salmonella typhimurium detection biosensor has the beneficial effects that the specific recognition of a nucleic acid aptamer is utilized; the high specific detection of salmonella typhimurium being a target object is realized by using the aptamer of salmonella typhimurium as a recognition substance; and nucleic acid tool enzymes are used for realizing the cyclic utilization of the target object and have a signal amplification effect.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to an electrochemical biosensor for detecting Salmonella typhimurium. Background technique [0002] Salmonella is a Gram-negative bacterium with blunt ends, no spores, and generally no capsule. Except for Salmonella pullorum and Salmonella gallinarum typhi, others have flagella all over the body. Salmonella is a common zoonotic pathogen that can not only cause gastroenteritis, but also various syndromes such as sepsis, typhoid fever, and extraintestinal focal infection. In particular, pathogenic bacteria represented by Salmonella typhimurium have caused a huge threat in food safety, environmental monitoring, disease prevention, etc., and have caused great harm to human health, so they have received strong attention from people. In recent years, the detection technology of Salmonella typhimurium has made great progress, from the very difficult traditional isolation and culture me...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/3277
Inventor 刘素裴倩倩黄加栋王玉王虹智郭玉娜
Owner UNIV OF JINAN
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