Method for portably and rapidly detecting ochratoxin A

Abstract

The invention discloses a method for portably and rapidly detecting ochratoxin A. The method comprises the following steps: first, preparing sucrose invertase-terminal alkyne modified DNA; subsequently, combining biotin-nitrine modified DNA with the surface of a magnetic bead, adding the ochratoxin A with different concentration, and mixing; adding the previously-prepared sucrose invertase-terminal alkyne modified DNA, a CuSO4 solution and an ascorbic acid solution, wherein the ascorbic acid solution can be used for reducing Cu(II) to obtain a Cu(I) compound so as to catalyze the biotin-nitrine modified DNA and the sucrose invertase-terminal alkyne modified DNA to have a click chemistry reaction; taking a supernatant liquor after the reaction is ended, then adding the supernatant liquor into a sucrose solution, wherein the sucrose invertase can effectively catalyze the sucrose in the solution to be transformed into glucose; and finally, measuring by adopting a glucometer. The method is used for organically combining the rapid reaction advantage of the click chemistry reaction with the high specificity characteristic of an aptamer, and adopting the simple and portable glucometer to acquire data, thus the operation complexity and detection cost are greatly reduced, and the sensitivity and selectivity are improved.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a portable and rapid detection method for ochratoxin A constructed based on a click chemical reaction. Background technique [0002] Ochratoxin A is one of the most common food contamination toxins, mainly produced by some Aspergillus and Penicillium fungi, which are easy to grow and multiply in cereal crops, fruits, beer and liquor. In animals, ochratoxin A is easy to be absorbed by the intestine, and it is easy to cause strong toxicity to the kidney and liver, and it also has carcinogenic, teratogenic, and immunosuppressive effects. The International Agency for Research on Cancer has listed ochratoxin A as a potential human carcinogen. The Food and Agriculture Organization of the United Nations / World Health Organization Joint Expert Committee on Food Additives (FAO / WHO JECFA) pointed out that 50% of ochratoxin A transmitted indirectly from food to humans comes f...

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Problems solved by technology

There have also emerged many methods to detect ochratoxin A, such as acceptable liquid chromatography, high performance liquid chromatography-mass spectrometry, and thin-layer chromatography, but these methods require expensive instruments and consume a lot of time. time, and requires specialized training personnel to operate, and the detection cost is high, thus hindering their application to a certain extent

At present, some highly sensitive immunoassay techniques have also been reported successively, such as enzyme-linked immunoassay, electrochemical immunosensor, fluorescent immunoassay technology, etc. However, these methods all need to rely on the preparation of antibodies, haptens, and immune antigens. Not only is it complicated and time-consuming, but it also needs to be tested with the help of large instruments, which is inconvenient to carry

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