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Biosensor for detecting salmonella typhimurium and application of biosensor

A biosensor, Salmonella technology, used in biological material analysis, instruments, measuring devices, etc., can solve the problems of high requirements for instruments and equipment, cumbersome detection methods, low sensitivity, etc. Effect

Active Publication Date: 2020-09-08
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The third object of the present invention is to prepare a magnetically separated quantum dot fluorescent sandwich sensor using DNA regular tetrahedral nucleic acid aptamer nanostructure as a signal recognizer and self-assembled carbon quantum dot microspheres as a signal amplifier, and using the sensor A method for detecting Salmonella typhimurium to solve the problems of low sensitivity, cumbersome detection methods, and high requirements for equipment and equipment in the existing detection technology for Salmonella typhimurium

Method used

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  • Biosensor for detecting salmonella typhimurium and application of biosensor
  • Biosensor for detecting salmonella typhimurium and application of biosensor
  • Biosensor for detecting salmonella typhimurium and application of biosensor

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Embodiment 1

[0059] The nucleic acid aptamer utilized in the present invention specifically acts on Salmonella typhimurium.

[0060] The sequence of Aptamer1 is: 5'-CTG ACAGAT TAAAAG TGG TTG GGC AAC TTC TGC TTGCGAA-3'

[0061] The sequence of the Aptamer2 is: 5'-TAT GGC GGC GTC ACC CGACGG GGA CTT GAC ATTATG ACA G-3'

[0062] A biosensor for detecting Salmonella typhimurium, which is composed of carbon quantum dot Td-CD modified by Salmonella typhimurium nucleic acid aptamer and magnetic beads Td-MB modified by Salmonella typhimurium nucleic acid aptamer;

[0063] Wherein, the Salmonella typhimurium aptamer is assembled with other nucleic acids through sequence design to form a DNA tetrahedral structure, and then used to modify carbon quantum dots and magnetic beads, and to modify the carbon quantum dots of Salmonella typhimurium. The DNA tetrahedral structure of the nucleic acid aptamer is Td-Aptamer1, and the DNA regular tetrahedral structure of the Salmonella typhimurium nucleic acid ap...

Embodiment 2

[0087] A method for detecting Salmonella typhimurium, the steps are:

[0088] 2.1 Standard curve drawing:

[0089]Take 6 1.5mL EP tubes, add 1mL of drinking water without Salmonella typhimurium to each, and add Salmonella typhimurium bacterial liquid to them so that the concentrations are 10CFU / mL, 10 2 CFU / mL, 10 3 CFU / mL, 10 4 CFU / mL, 10 5 CFU / mL, 10 6 CFU / mL. Take another 6 1.5mL EP tubes, add 100μL of bacterial solution, 1mL of Td-MB solution and 1mL of Td-CD solution and incubate at room temperature for 20 minutes, shake slowly to fully combine the reaction; separate on a magnetic rack, discard the supernatant to obtain MB-Salmonella typhimurium-CD complex; use 1.5mL PBS solution to resuspend the complex, then add a certain concentration of Biotin molecular solution 2mL and mix for 30 minutes; then, perform the second separation of magnetic beads, discard the magnetic beads, and keep Supernatant solution; Finally, the solution after the reaction is scanned by a fluo...

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Abstract

The invention discloses a biosensor for detecting salmonella typhimurium and application of the biosensor, and belongs to the technical field of harmful microorganism detection. The biosensor is composed of carbon quantum dots Td-CD modified by a salmonella typhimurium nucleic acid aptamer and magnetic beads Td-MB modified by the salmonella typhimurium nucleic acid aptamer. The salmonella typhimurium nucleic acid aptamer and other nucleic acids are assembled through sequence design to form a DNA regular tetrahedron structure, and then the DNA regular tetrahedron structure is respectively usedfor modifying carbon quantum dots and magnetic beads. The method for detecting salmonella typhimurium by adopting the biosensor is simple and convenient to operate and short in consumed time, complexpretreatment does not need to be carried out on a sample to be detected, and the detection cost is low; the biosensor can effectively amplify signals of salmonella typhimurium, can be used for detecting low-concentration salmonella typhimurium in food, and achieves qualitative and quantitative analysis of salmonella typhimurium.

Description

technical field [0001] The invention belongs to the technical field of harmful microorganism detection, and in particular relates to a biosensor for detecting Salmonella typhimurium and an application thereof. Background technique [0002] Salmonella typhimurium (Salmonella typhimurium) is a common food-borne pathogen. It is a Gram-negative anaerobic bacterium that widely exists in food, water sources, animals and plants. Contaminated food and water can cause mammals and Human gastroenteritis. Common symptoms caused by Salmonella typhimurium infection include high fever, cough, diarrhea, abdominal cramps and vomiting. Severe illness can lead to death, causing huge losses to food safety and human health. Therefore, it is particularly important to accurately identify and detect Salmonella typhimurium. [0003] At present, the methods for detecting Salmonella typhimurium include biological isolation and culture method, enzyme-linked immunoassay method, PCR molecular detection...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N21/64
CPCG01N33/56916G01N21/6428G01N2333/255Y02A50/30
Inventor 吴薇杨庆利都晗
Owner QINGDAO AGRI UNIV
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