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Rapid detection method of alternariol in wheat flour based on nucleic acid adaptation

A technology for the detection of alternariol and its detection method, which is applied in the field of rapid detection of alternariol in wheat flour, which can solve the problems of expensive equipment, long detection time, time-consuming and laborious, etc., and achieve short synthesis cycle, low cost and improved stability and accuracy effects

Inactive Publication Date: 2021-01-08
浙江科德生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these traditional detection methods usually require large-scale, expensive equipment and professional technicians, and the detection can only be carried out in the laboratory, which takes a long time and is time-consuming and laborious.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Synthesize DNAzyme 1 and its complementary DNAzyme 2.

[0036] The DNAzyme aptamer DNAzyme 1 sequence is 5'SH-ATACTCAGGTACGCTTGATCGATCGATCGCTAGATGCAT-3'.

[0037] The sequence of DNAzyme 2 complementary strand is 5'SH-TATGAGTCCATGCGAACTAGCTAGCTAGCGATCTACGTA-3'.

[0038] 2. Preparation of gold nanoparticles.

[0039](1) Dilute 1ml of 1% HAuCl4 solution to 100ml, heat to boiling, after 10min, quickly add 4ml of 1% trisodium citrate aqueous solution, continue to boil for 10min to appear transparent orange-red.

[0040] (2) Stop heating, continue to stir for 15 minutes, cool to room temperature, filter, and dry at 50°C for 6 hours.

[0041] 3. Preparation of DNAzyme 1-nanogold probes.

[0042] (1) Take a 2ml centrifuge tube, and dissolve 5mg of the nano-gold particles prepared in the previous step in 1ml of dd water to make a 0.5% nano-gold solution.

[0043] (2) Put 50 μl of 10 μmol / L deoxyribozyme aptamer DNAzyme 1 into a 1.5ml centrifuge tube, add 40 μl of 10 mmol...

Embodiment 2

[0059] 1. The preparation method of aptamer-nano-gold probe and complementary strand nano-probe is the same as that in Example 1.

[0060] 2. Detection of Alternariol.

[0061] (1) Take 10g of wheat, grind it, add 2ml of dd water, oscillate and filter evenly, and take the filtrate as the solution to be tested for later use.

[0062] (2) Take 100 μl DNAzyme 1-nano-gold probe, 100 μl DNAzyme 2-nano-gold probe, 100 μl test solution and 50 μl PB and buffer, put them in a 1.5ml centrifuge tube, and incubate at 37°C for 1 hour.

[0063] (2) Add 17.25μl 2mol / L NaCl solution to make the NaCl concentration reach 100mmol / L, observe the color change of the solution, and judge the content and approximate content of alternariol in wheat according to the depth of the solution color.

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Abstract

The invention discloses a rapid detection method of alternariol in wheat flour based on nucleic acid adaptation. The method utilizes an aptamer probe to specifically capture alternariol, thereby effectively enhancing the detection stability and accuracy. The alternariol is a small molecular substance, forms a competitive relationship with the nanogold after being combined with the aptamer, can bedistinguished by naked eyes by observing the color change of the solution based on the agglomeration condition of the nanogold; and using of a complex instrument is not needed, and on-site real-time detection is realized. The aptamer probe used in the method has the advantages of being capable of being artificially synthesized, convenient to modify, short in synthesis period, low in cost, good instability, small in batch-to-batch difference and capable of being stored for a long time, and can be stored after mass production, so that the detection period can be shortened. Meanwhile, visual rapid detection and semi-quantitative detection of the alternariol in the wheat can be realized according to the color change degree of the solution.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid detection method for alternariol in wheat flour. Background technique [0002] Alternaria toxins are common saprophytic bacteria in soil, air, and industrial materials, and are mainly a series of toxic secondary metabolites produced by Alternaria spp. Because Alternaria can grow and reproduce at low temperature, it is an important cause of spoilage of agricultural products during storage and transportation. At present, there are more than 70 species known to have obvious toxicity, but few of them have been reported and confirmed. Among them, 10 species have toxic effects on animals and plants. Alternariol (AOH) and Alternariol monomethyl ether (AME) belong to dibenzopyrone toxins, which are mutagenic, genotoxic and carcinogenic, etc. Toxic small molecule compounds. The results of two risk assessments conducted by EFSA in 2011 and 2016 indicated that dietary intake of AOH a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/25G01N21/78
CPCG01N21/25G01N21/78G01N2021/258
Inventor 娄新徽翁国武朱沉雷
Owner 浙江科德生物科技有限公司
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