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Biosensor for detection of Salmonella typhimurium based on nucleic acid aptamer and preparation method thereof

A biosensor and nucleic acid aptamer technology, applied in the field of biosensors, can solve the problems of high cost, low specificity and sensitivity, etc., and achieve the effects of low detection limit, simple preparation method, and accurate quantitative detection

Inactive Publication Date: 2018-09-04
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of relatively low specificity and sensitivity and high cost of the method for detecting Salmonella typhimurium in the above prior art, the present invention provides a nucleic acid aptamer-based nucleic acid aptamer with high specificity and sensitivity, low cost and fast detection speed. Biosensor for detection of Salmonella typhimurium

Method used

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  • Biosensor for detection of Salmonella typhimurium based on nucleic acid aptamer and preparation method thereof
  • Biosensor for detection of Salmonella typhimurium based on nucleic acid aptamer and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0050] The main steps of the electrode modification process are as follows:

[0051] a. The gold electrode is first polished in 0.3 μm and 0.05 μm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0052] b. Add 10 μL of HAP2 (10 μM) dropwise to the surface of the electrode, and incubate at room temperature (37° C.) for 2 h. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0053] 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:

[0054] a. Add sterilized water, Aptamer (1 μL), 1 μL of Primer chains of different concentrations (concentrations are 20 μM, 10 μM, 5 μM, 2 μM, 1 μM) and the target substance to the pre-prepared sterilized No. 1-5 centrifuge tube. Shake for 30s, then put the mixed mixture into a 37°C incubator and incubate for 1h;

[0055] b. Add 10× b...

Embodiment 2

[0067] A preparation method of the electrochemical biosensor of the present invention:

[0068] The main steps of the electrode modification process are as follows:

[0069] a. The gold electrode is first polished in 0.3 μm and 0.05 μm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0070] b. Add 10 μL of HAP2 (10 μM) dropwise to the surface of the electrode, and incubate at room temperature for 2 hours. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0071] 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:

[0072] a. Add sterilized water, Primer (1 μL), 1 μL of different concentrations of Aptamer chains (concentrations are 20 μM, 10 μM, 5 μM, 2 μM, 1 μM) and the target substance to the pre-prepared sterilized No. 1-5 centrifuge tube. Shake for 30s, then pu...

Embodiment 3

[0080] A preparation method of the electrochemical biosensor of the present invention:

[0081] The main steps of the electrode modification process are as follows:

[0082] a. The gold electrode is first polished in 0.3 μm and 0.05 μm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0083] b. Add 10 μL of HAP2 (10 μM) dropwise to the surface of the electrode, and incubate at room temperature for 1 h. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0084] 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:

[0085] a. Add sterilized water, Primer (1 μL), Aptamer chain (1 μL) and the target substance into pre-prepared sterilized centrifuge tubes No. 1-5. Shake for 30s, then put the mixed mixture into a 37°C incubator and incubate for 1h;

[0086] b. Add 10× buff...

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Abstract

The invention relates to an aptamer based salmonella typhimurium detection biosensor and a preparation method thereof. Signal amplification is realized by continuous generation and cyclic utilization of Primer, and consequently high-sensitivity detection of salmonella typhimurium is realized, and lower detection limit can be acquired. The aptamer based salmonella typhimurium detection biosensor is high in specificity and sensitivity, low in cost and quick in detection. By the detection method, a salmonella typhimurium detection limit is 5-15cfu / mL, and a detection range is 1*10-1*107cfu / mL.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a biosensor for detecting Salmonella typhimurium based on nucleic acid aptamers, and also relates to a preparation method thereof. Background technique [0002] Salmonella is one of the important food-borne pathogens of Enterobacteriaceae, which accounts for a relatively high proportion of food poisoning cases in my country. Salmonella typhimurium has been widely concerned because of its strong drug resistance, wide transmission route, and multiple host infections. Studies have shown that Salmonella typhimurium has extremely strong multi-drug resistance, showing resistance to most antibiotics. In the survey, the number of Salmonella typhimurium infections accounted for 17% of the total Salmonella infections that year. At the same time, Salmonella typhimurium is also one of the most common serotypes among more than 290 serotypes of Salmonella found in my country. [0003] At...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6816C12Q1/04
CPCC12Q1/6825C12Q2521/301C12Q2525/205Y02A50/30
Inventor 刘素王玉邱婷婷黄加栋王虹智郭玉娜崔洁许颖崔雪君裴倩倩冷雪琪韩聪
Owner UNIV OF JINAN
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