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Production method of novel fluorescent biosensor for bacillus coli detection

A biosensor, Escherichia coli technology, applied in the directions of fluorescence/phosphorescence, instruments, measuring devices, etc., can solve the problems of early diagnosis of unfavorable diseases, low detection limit, unfavorable promotion, etc., and achieve significant technological innovation significance and commercial value, and the method is simple , the effect of expanding the range

Inactive Publication Date: 2009-07-01
HUNAN UNIV OF TECH +3
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Problems solved by technology

Although the immunoassay has high specificity, the detection limit is low, often only reaching 10 3 ~10 8 CFU / ml, and it needs two times of enrichment, which takes a long time and is not conducive to the early diagnosis of diseases
Although molecular biology detection technology has high sensitivity, especially the latest DNA probe technology or gene chip technology can reach 10 2 ~10 3 The lowest detection limit of CFU / ml or even dozens of CFU / ml, but there are often some false positives, and it requires large-scale equipment, professionals and personnel, which is not conducive to the promotion in grassroots medical institutions and environmental monitoring

Method used

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  • Production method of novel fluorescent biosensor for bacillus coli detection
  • Production method of novel fluorescent biosensor for bacillus coli detection
  • Production method of novel fluorescent biosensor for bacillus coli detection

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Experimental program
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Embodiment approach I

[0023] figure 1 It represents one of the preparation processes of the solid-phase microarray: the preparation method of the fluorescent biosensor microarray by the magnetic particle in situ method. The steps and principles are briefly described as follows: 1) The surface of the solid-phase microarrayed magnetic particles is modified with a secondary antibody, and the primary antibody is labeled with a fluorescent probe (such as fluorescein), and then the fluorescently labeled primary antibody and the secondary antibody are mixed. Combine to form an antibody complex; 2) use the prepared antibody complex to process the E. coli O157:H7 antigen sample to obtain a negatively charged E. coli O157:H7 antigen-antibody conjugate; 3) negatively charged antigen on the magnetic particles After the antibody conjugate is specifically combined with the cationic conjugated polyelectrolyte (CCP), a supramolecular recognition system is formed, and the light excitation of the CCP leads to resona...

Embodiment approach II

[0025] figure 2 It represents the second preparation process of the solid-phase microarray: the preparation method of the fluorescent biosensor microarray by the shear spotting method. The steps and principles are briefly explained as follows: 1) The surface of the magnetic particle is biofunctionalized to modify the secondary antibody, and the primary antibody is labeled with a fluorescent probe (such as fluorescein), and the labeled primary antibody binds to the secondary antibody to form an antibody complex; 2) Treat Escherichia coli O157:H7 antigen samples with antibody complexes, and then obtain negatively charged Escherichia coli O157:H7 antigen-antibody conjugates through shearing and magnetic separation processes; 3) Negatively charged Escherichia coli O157:H7 antigen-antibody conjugates After the conjugate is spotted and specifically combined with the cationic conjugated polyelectrolyte (CCP), a supramolecular recognition system microarray is formed, and the CCP ligh...

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Abstract

The invention relates to a super sensitivity fluorescence biosensor, for using magnetic practical to load antibodies and capture antigens, using water soluble conjugated polyelectrolyte to induce fluorescence amplification to check Escherichia coli O157: H7, which first utilizes magnetic particles to capture Escherichia coli O157: H7 antigens, to generate an antigen antibody unity with fluorescence labels; adds water soluble conjugated polyelectrolyte to induce and act with electrostatics, to recognize the super-molecules between the antigen antibody unity and the conjugated polyelectrolyte to form a super-molecule fluorescence composite system; exciting the fluorescence composite via the light or electricity of the conjugated polyelectrolyte, to induce the super fluorescence amplification of thousands and millions level of the prior fluorescence label signals, thereby forming a super sensitivity fluorescence biosensor. The biosensor system is based on the fluorescence sensitivity raising and efficiency raising effect of water soluble conjugated polyelectrolyte, which can be applied for fast super sensitivity detection on acute infectious disease as Escherichia coli O157: H7.

Description

Technical field [0001] The invention relates to a method for preparing a novel fluorescent biosensor of ultrasensitive and highly selective Escherichia coli O157:H7 induced by conjugated polyelectrolyte to amplify fluorescence. Background technique [0002] With the penetration of nanotechnology in the field of biosensors, the two most advanced and most researched types in the field of biosensors are electrochemical biosensors and optical biosensors. Electrochemical biosensors have the advantages of high sensitivity, easy miniaturization, and the ability to operate in turbid solutions, and the required instruments are simple and cheap. Therefore, it is widely used in the preparation of biosensors; in comparison, the advantages of optical biosensors are that they do not require a reference electrode, are not disturbed by electromagnetic fields, can measure certain substances (such as oxygen) under non-equilibrium conditions, and have good stability. , especially in the case ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/536G01N21/64
Inventor 贺全国黄景科曾晓希
Owner HUNAN UNIV OF TECH
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