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Ultrasensitive colorimetric sensing detection method for determining ochratoxin A

A technology for ochratoxin and colorimetric sensing, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., which can solve the problems that limit the large-scale application of such methods, limit the sensitivity and accuracy of immunological methods, and expensive equipment. And other issues

Inactive Publication Date: 2018-02-23
樊之雄
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0006] Technical problems to be solved: Due to the expensive equipment and time-consuming sample preparation of traditional instrument detection methods, it is not suitable for the screening of a large number of samples, which limits the large-scale application of such methods; immunological methods based on antigen antibodies rely on the sensitivity and specificity of antibodies, Therefore, the quality of the antibody limits the sensitivity and accuracy of the immunological method

Method used

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  • Ultrasensitive colorimetric sensing detection method for determining ochratoxin A
  • Ultrasensitive colorimetric sensing detection method for determining ochratoxin A
  • Ultrasensitive colorimetric sensing detection method for determining ochratoxin A

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Embodiment 1

[0036] An ultrasensitive colorimetric sensing detection method for measuring ochratoxin A, comprising the following specific steps:

[0037] (1) Magnetic nanoparticles modified ochratoxin A aptamer

[0038] The newly synthesized particle size is 300nm concentration is 1mg mL -1 The carboxyl-modified magnetic nanoparticles were diluted 10 times with 0.01M phosphate buffer at pH 7.4, and then 0.068 mg carbodiimide and 0.15 mg N-hydroxysulfosuccinate were added to 1 mL of the diluted magnetic nanoparticles imide, put it on a shaker and activate it by shaking at room temperature for 20 min, then add 50 μL of 3’-terminal amino-modified ochratoxin A aptamer DNA fragment with a concentration of 10 μM to the above activated magnetic nanoparticles, After shaking and reacting for 3 hours at room temperature, use a magnetic frame to absorb the magnetic nanoparticles to aggregate at the bottom of the centrifuge tube, and remove the supernatant to remove uncoupled free aptamer DNA molecul...

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Abstract

The invention relates to an ultrasensitive colorimetric sensing detection method for determining ochratoxin A, belonging to the field of biological detection. The method comprises the steps of modifying an ochratoxin A aptamer by virtue of magnetic nanoparticles, forming a partially hybridized DNA double-chain structure by virtue of the magnetic nanoparticles modified aptamer and partially complementary DNA molecules, preparing a DNA modified gold nanoparticle probe, and constructing an ochratoxin A colorimetric sensor. According to the method, by virtue of a specific recognition function of the ochratoxin A aptamer to ochratoxin A molecules, and the assembling degrees of gold nanoparticles are differentiated through ligase chain reaction when ochratoxins A with different concentrations exist, so that different color gradients are formed, and the target ochratoxin A is detected by determining the change of an ultraviolet absorption value after the reaction. The method is high in detection sensitivity, low in detection limit and good in specificity, a detection method is easy to observe, and the effective method is provided for the trace detection of the ochratoxin A.

Description

technical field [0001] The invention discloses an ultrasensitive colorimetric sensing detection method for determining ochratoxin A, which belongs to the field of biological detection. Background technique [0002] Ochratoxin is a group of structural analogs produced by fungi, mainly including four compounds A, B, C, and D, among which the toxicity is the most, the most closely related to human health, the highest toxin production, the most serious pollution to crops, and the distribution The most widespread is ochratoxin A. Ochratoxin A is a toxic metabolite produced by Aspergillus and Penicillium, mainly produced by Penicillium puree, Ochratoxin and Aspergillus carbon niger, widely distributed in nature, in grains and their products, animal feed, animal products, Products such as cocoa and chocolate have a high pollution rate, have strong kidney toxicity and liver toxicity, and have teratogenic, mutagenic and carcinogenic effects, so they pose a huge potential threat to h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6862
CPCC12Q1/6862C12Q2525/205C12Q2563/143C12Q2563/155C12Q2563/137
Inventor 樊之雄
Owner 樊之雄
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