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Nucleic acid aptamer-quantum dot biosensor for detecting and killing salmonella and preparation method and application thereof

A Salmonella nucleic acid and nucleic acid aptamer technology, which is applied in measurement devices, biological testing, biological material analysis, etc., can solve the problems of high requirements for instruments and equipment, cumbersome detection methods, incomplete sterilization, etc., and achieves high sensitivity and significant sterilization effect. , use a wide range of effects

Active Publication Date: 2021-06-01
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned biosensor and sterilization method of the present invention can solve the problems of low sensitivity, cumbersome detection methods, high requirements for instruments and equipment, and incomplete sterilization in the existing technology for detecting and killing Salmonella

Method used

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  • Nucleic acid aptamer-quantum dot biosensor for detecting and killing salmonella and preparation method and application thereof
  • Nucleic acid aptamer-quantum dot biosensor for detecting and killing salmonella and preparation method and application thereof
  • Nucleic acid aptamer-quantum dot biosensor for detecting and killing salmonella and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0045] The nucleic acid aptamer-quantum dot biosensor for the detection and elimination of Salmonella was deposited on a PDA-coated Fe 3 o 4 The surface of the nanoparticle is connected with the Salmonella nucleic acid aptamer Aptamer C (SEQ ID NO.1) labeled with carbon quantum dots; and the complementary chain Strand A (SEQ ID NO.2) modified with fluorescein FAM is paired with the above-mentioned Aptamer C;

[0046] The 5' of the Salmonella nucleic acid aptamer Aptamer C is modified with a carboxyl group, and the 3' is modified with -SH, specifically as follows:

[0047] 5'-COOH-AGCAGCACAG-GCGATCCAAGCTTCTTCA-SH-3'

[0048] The complementary strand Strand A modified with fluorescein FAM is as follows:

[0049] 3'-CGCTAGGTTCGAAGAAGT-FAM-5'

[0050] The carbon quantum dots use ascorbic acid as a carbon source, and the carbon quantum dots with a particle size of 2-3nm and good fluorescence properties are prepared in one step by a hydrothermal method.

[0051] The above-mentio...

Embodiment 2

[0059]A method for detecting and disinfecting salmonella, the steps are as follows:

[0060] The nucleic acid aptamer-quantum dot biosensor of Example 1 was dispersed in phosphate-buffered saline to prepare a solution with a concentration of 0.32 μM, and the above-mentioned dispersion was added to the surface of the food for detection of Salmonella in it. The effect is specifically combined with the Aptamer C chain, and the A chain will be competed by bacteria, and the carbon quantum dots on the Aptamer C chain will show blue-green fluorescence due to the loss of the quenching effect of the fluorescein FAM on the A chain. It is used to determine the content of Salmonella by detecting the blue-green fluorescence of carbon quantum dots;

[0061] In the area showing fluorescence, use 1.5W / cm 2 808nm near-infrared light is continuously irradiated; after the sterilization is completed, the food is placed in a magnetic field or Fe is separated under the action of a magnet 3 o 4 ...

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Abstract

The invention discloses a nucleic acid aptamer-quantum dot biosensor for detecting and killing salmonella and a preparation method and application thereof, and belongs to the technical field of detection and inactivation of food pathogenic bacteria. The biosensor disclosed by the invention is prepared by connecting a salmonella aptamer Aptamer C marked by quantum dots to the surfaces of Fe3O4 nanoparticles deposited with a polydopamine (PDA) coating; pairing a complementary chain Strand A modified with fluorescein with the Aptamer C, and pairing a complementary chain Strand A modified with fluorescein with the Aptamer C; using the Fe3O4 nano particles connected with the Aptamer C and the Srand A for detecting the salmonella. The detection sensitivity of the biosensor is 10 CFU / 100 g, the detection range is 10-10< 4 > CFU / 100 g, and the biosensor has good sensitivity and specificity; the sterilization degree reaches 95% or above, and the effect is remarkable.

Description

technical field [0001] The invention belongs to the technical field of detection and deactivation of food pathogenic bacteria, and in particular relates to a nucleic acid aptamer-quantum dot biosensor for detection and deactivation of Salmonella, its preparation method and application. Background technique [0002] Food pathogens are pathogenic bacteria that can cause food poisoning or use food as a medium of transmission. Pathogenic bacteria directly or indirectly contaminate food and water sources, and human oral infection can lead to the prevalence of intestinal infectious diseases, food poisoning, and livestock and poultry infectious diseases. Therefore, foodborne pathogens are an important source of food safety problems. [0003] The impact of food-borne pathogens on human health: (1) Acute poisoning: In general, food-borne pathogens often cause acute poisoning, and the milder ones often appear as symptoms of acute gastroenteritis, such as vomiting, nausea, Abdominal ...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/569G01N33/58G01N33/543A23L5/20A23B4/01
CPCG01N21/6428G01N33/56916G01N33/588G01N33/54346A23L5/21A23B4/01G01N2021/6432G01N2021/6439G01N2333/255A23V2002/00A23V2300/24A23V2200/10Y02A50/30
Inventor 吴薇孙霏霏杨庆利唐娟
Owner QINGDAO AGRI UNIV
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