Rapid detection method for ochratoxin A

A technology for ochratoxin and detection method, which is applied in the field of rapid detection of ochratoxin A, and can solve the problems of complex, time-consuming, expensive instruments and the like

Active Publication Date: 2017-05-24
武汉市农业科学技术研究院农业环境安全检测研究所
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Problems solved by technology

Although these methods have high sensitivity and high accuracy, they all require

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Embodiment 1

[0022] The present embodiment provides a kind of OTA rapid detection method, comprises the following steps:

[0023] (1) Add 10mg of aminated microporous silicon nanomaterials to 2mL containing 30mg mL -1 In the Tris-HCl buffer solution (10mM, pH 7.0) of methylene blue, slightly shake overnight at room temperature, so that the methylene blue molecules can fully diffuse into the pores of the microporous silicon nanomaterial;

[0024] (2) Add 20 μL nucleic acid aptamer solution (200 μM) to the suspension obtained in step (1), stir gently at 4 °C for 8 h, centrifuge and wash with Tris-HCl buffer (10 mM, pH 7.0) for more than 3 times ( To remove the methylene blue physically adsorbed on the surface of the microporous silicon nanomaterial), and finally the material is dispersed in 2mL Tris-HCl buffer solution;

[0025] (3) Add 400 μL of the mixture of OTA sample and nuclease (choose more than 10 concentrations from 0 to 2000 nM) to 400 μL of the suspension prepared in step (2), an...

Embodiment 2

[0028] The present embodiment provides a kind of OTA rapid detection method, comprises the following steps:

[0029] (1) Add 5 mg of positively charged microporous silicon nanomaterials on the surface to 500 mL of Tris-HCl buffer solution (pH 7.5) containing 2.0 M glucose, and shake slightly overnight at room temperature to fully diffuse the glucose molecules into the microporous silicon nanomaterials. in the hole;

[0030] (2) Add 20 μL nucleic acid aptamer solution (200 μM) to the above suspension, stir gently at 4°C for 8 hours, centrifuge and wash with Tris-HCl buffer (10 mM, pH 7.5) for more than 3 times (to remove microbes The methylene blue physically adsorbed on the surface of porous silicon nanomaterials), and finally the material was dispersed in 2mL Tris-HCl buffer (C [微孔硅] ≈25mg mL -1 );

[0031] (3) Add 400 μL of a mixture of OTA samples and nucleases of different concentrations (select more than 10 concentrations from 0 to 3000 nM) to 400 μL of the suspension ...

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Abstract

The invention provides a rapid detection method for ochratoxin A. The method comprises the following steps that a large number of methylene blue molecules are loaded in a microporous silicon nano-material; an aptamer of the ochratoxin A closes the microporous silicon nano-material loaded with the methylene blue molecules; after ochratoxin A samples are added, due to the fact that the ochratoxin A samples and the aptamer are subjected to specific binding, the aptamer comes away from the microporous silicon nano-material surface, and the methylene blue molecules are released. By means of screen-printed electrodes and a portable electrochemical detector, an electric signal of methylene blue in supernatant is detected, a relation curve between the electric signal and the concentration of the ochratoxin A, and therefore indirect quantitative detection of the ochratoxin A is achieved. On the basis, the signal amplification effect of an enzyme is introduced. After nuclease is added, a G-quadruplex structure formed between the ochratoxin A and the aptamer is selectively destroyed, the ochratoxin A returns to a solution in a free state, the above mentioned processes are repeated. In this way, the sensitivity of detection can be greatly improved. According to the method, the sensor building process and detection process do not need expensive instruments and reagents, operation is easy, and the sensitivity is very high.

Description

technical field [0001] The invention relates to a rapid detection method for ochratoxin A, in particular to a rapid detection method for ochratoxin A based on microporous silicon nanomaterials, nuclease cycle signal amplification and screen-printed electrodes. Background technique [0002] Ochratoxins (ochratoxins) are a class of compounds produced by a variety of Aspergillus and Penicillium, which are called ochratoxin A (OTA), ochratoxin B (OTB) and ochratoxin C (OTC) in the order of their discovery. . It is another mycotoxin that has attracted worldwide attention after aflatoxin. Among them, OTA is the most toxic, the most widely distributed, the highest toxin production, the most polluting to agricultural products, and the most closely related to human health. It is a colorless crystalline compound, soluble in polarity Organic solvent and dilute sodium bicarbonate solution, slightly soluble in water, has a high chemical stability and thermal stability. This toxin i...

Claims

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Application Information

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IPC IPC(8): G01N27/30G01N27/26
CPCG01N27/26G01N27/30
Inventor 王利华曾令文王梦韩艳云杨保国吴文辉胡志娟
Owner 武汉市农业科学技术研究院农业环境安全检测研究所
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