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Fluorescent probe and preparation method thereof, biosensor as well as building method and application thereof

A biosensor and fluorescent probe technology, applied in the field of biosensing, can solve the problems of inaccurate detection results of markers, susceptibility to environmental interference, and unsatisfactory sensitivity, and achieve the goal of eliminating built-in interference, reducing detection limit, and improving sensitivity Effect

Inactive Publication Date: 2019-10-18
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The traditional methods for detecting the concentration of 2,6-pyridinedicarboxylic acid mainly include fluorescent polymerase chain reaction detection method, colorimetric method, high performance liquid chromatography, etc. Although these methods have reliable detection results in a single system, the processing process Complicated, high detection limit, small detection range, and inaccurate detection results for markers in complex systems, poor selectivity, unsatisfactory sensitivity, and easy to be affected by environmental interference

Method used

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  • Fluorescent probe and preparation method thereof, biosensor as well as building method and application thereof
  • Fluorescent probe and preparation method thereof, biosensor as well as building method and application thereof
  • Fluorescent probe and preparation method thereof, biosensor as well as building method and application thereof

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Embodiment 1

[0041] A biosensor for detecting the concentration of 2,6-pyridinedicarboxylic acid by fluorescence method of the present invention, comprising a fluorescent probe of europium-doped gold nanocluster and a standard curve of standard 2,6-pyridinedicarboxylic acid concentration; europium The fluorescent probe doped with the gold nanocluster is obtained by doping the gold nanocluster with europium ions, and the particle size of the fluorescent probe is 1-2 nm.

[0042] The biosensor construction method comprises the steps of:

[0043] (1) Synthesis of fluorescent probes by one-pot method: prepare 8mM chloroauric acid solution and europium nitrate solution respectively, mix them uniformly at a volume ratio of 1:1, and stir vigorously to obtain a reaction containing europium elements system;

[0044] (2) Add reducing agent: add bovine serum albumin as reducing agent and stabilizer in the reaction system, its concentration is 70mg / mL, stir vigorously at the speed of 1000r / min during...

Embodiment 2

[0051] A biosensor for detecting the concentration of 2,6-pyridinedicarboxylic acid by fluorescence method of the present invention, comprising a fluorescent probe of europium-doped gold nanocluster and a standard curve of standard 2,6-pyridinedicarboxylic acid concentration; europium The fluorescent probe doped with the gold nanocluster is obtained by doping the gold nanocluster with europium ions, and the particle size of the fluorescent probe is 1-2 nm.

[0052] The biosensor construction method comprises the steps of:

[0053] (1) Fluorescent probes were synthesized by a one-pot method: 5mM chloroauric acid solution and 10mM europium nitrate solution were prepared respectively, and the two were mixed uniformly at a volume ratio of 1:1, and vigorously stirred to obtain europium-containing elements reaction system;

[0054] (2) Adding reducing agent: Bovine serum albumin was added as reducing agent and stabilizer in the reaction system, its concentration was 50 mg / mL, and i...

Embodiment 3

[0061] A biosensor for detecting the concentration of 2,6-pyridinedicarboxylic acid by fluorescence method of the present invention, comprising a fluorescent probe of europium-doped gold nanocluster and a standard curve of standard 2,6-pyridinedicarboxylic acid concentration; europium The fluorescent probe doped with the gold nanocluster is obtained by doping the gold nanocluster with europium ions, and the particle size of the fluorescent probe is 1-2 nm.

[0062] The biosensor construction method comprises the steps of:

[0063] (1) Synthesis of fluorescent probes by one-pot method: Prepare 10mM chloroauric acid solution and europium nitrate solution respectively, mix them uniformly at a volume ratio of 1:1, and stir vigorously to obtain a reaction containing europium elements system;

[0064] (2) Add reducing agent: add bovine serum albumin as reducing agent and stabilizer in the reaction system, its concentration is 100mg / mL, stir vigorously at the speed of 1000r / min duri...

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Abstract

The invention discloses a fluorescent probe for detecting the concentration of 2,6-dipicolinic acid with a fluorescent method. The fluorescent probe comprises gold nano-cluster particles doped with europium elements, and the particle size of the fluorescent probe is 1 to 2nm. Through combination of the luminescence of rare-earth europium elements and the stable luminescence of gold nano-clusters,the obtained europium-doped gold nano-cluster fluorescent probe has the characteristic of ratio-dependent luminescence, so that the environmental built-in interference is eliminated; a stable luminescence signal, high detection accuracy, high sensitivity and a wide detection range are achieved; and the detection limit is lowered. The invention also provides a biosensor for detecting the concentration of 2,6-dipicolinic acid with the fluorescent method. Through combination of the europium-doped gold nano-cluster fluorescent probe and a standard curve, subsequent detection steps can be simplified; high sensitivity, high accuracy and a wide detection range are achieved; and other precise and precious instruments are not required. The invention also discloses preparation methods of the fluorescent probe and the biosensor. The preparation methods have the advantages of simple preparation process, easiness in operation, low cost and suitability for popularization and use.

Description

technical field [0001] The invention belongs to the technical field of biosensing, and in particular relates to a fluorescent probe for detecting 2,6-pyridinedicarboxylic acid by a fluorescence method, a preparation method thereof, a biosensor, a construction method and application thereof. Background technique [0002] 2,6-pyridinedicarboxylic acid is a nuclear substance released during the germination of a dormant spore of a food-borne marker pathogenic bacteria, and it exists in large quantities in bacterial spores. The traditional heat sterilization technology can only kill the bacterial vegetative body, but not the spores. The strong stress resistance of spores mainly depends on the highly dehydrated state of the spore core area or its inner cytoplasm and the high density of nuclear substances such as calcium dipicolinate. During the germination process of spores, 2,6-pyridinedicarboxylic acid and its combined divalent cations (mainly Ca 2+ ) are released from the spo...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/64G01N21/6428G01N2021/6421
Inventor 阳明辉李晓庆候轶
Owner CENT SOUTH UNIV
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