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Method for signal amplification fluorescence detection of aflatoxin B1 based on hybridization chain reaction and DNAzyme

A signal amplification and aflatoxin technology, which is applied in the fields of analytical chemistry and biosensing detection, can solve the problems of troublesome pretreatment of graphene oxide, harsh environment required for the reaction system, and failure to achieve signal amplification, etc., to achieve high Sensitivity and selectivity, high actual sample detection ability, effect of improving detection sensitivity

Inactive Publication Date: 2018-04-24
OCEAN UNIV OF CHINA
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Problems solved by technology

However, the above methods also have certain shortcomings. The strategy of Goud and Seok did not achieve signal amplification; although Zhang achieved signal amplification, the pretreatment of graphene oxide was more troublesome; The environment required by the system is relatively harsh

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  • Method for signal amplification fluorescence detection of aflatoxin B1 based on hybridization chain reaction and DNAzyme
  • Method for signal amplification fluorescence detection of aflatoxin B1 based on hybridization chain reaction and DNAzyme
  • Method for signal amplification fluorescence detection of aflatoxin B1 based on hybridization chain reaction and DNAzyme

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Embodiment 1

[0027] Example 1: Establishment of a method for detecting aflatoxin B1 based on hybridization chain reaction and DNAzyme signal amplification and fluorescence detection

[0028] 1. Reaction time optimization for chain hybridization

[0029] figure 2 It is the corresponding fluorescence signal spectrum at different reaction times after adding the hairpin probe. It can be seen from the figure that within 2 to 6 hours, the fluorescent signal becomes stronger with the increase of the hybridization chain reaction time. When the reaction time is 7 hours, the signal is almost the same as that of 6 hours. Therefore, in the experiment We choose the reaction time of chain hybridization to be 6 hours.

[0030] 2. Fluorescence sensor sensitivity

[0031] The effect of reaction time on fluorescence intensity was investigated systematically. The results show that the addition of Mg to the system 2+ Finally, when the reaction time was 40min, the fluorescence signal was the strongest. ...

Embodiment 2

[0034] Embodiment 2: detection of aflatoxin B1 in moldy samples

[0035] After crushing 2 g of rice, add 6 mL of methanol / water solution (methanol:water=8:2), vortex for 30 min, centrifuge at 5000 rpm for 10 min, take the supernatant and filter it with a 0.45 μM filter head, and add different concentrations of The standard was made into the solution to be tested. Subsequently, detection was carried out according to the optimized conditions. Detection Table 2 is the detection data of aflatoxin B1 in the sample extract. From the data in Table 2 combined with the calculation of the concentration of the sample solution, it can be seen that the aflatoxin content is about 9.24±1.34 ng / g and 7.82±0.62 ng / g, which has exceeded the European standard of 2 μg / kg and is close to the national standard of 10 μg / kg, indicating that Moldy samples contain a certain amount of aflatoxin B1 and are not edible.

[0036] Table 2 Determination of aflatoxin B1 content in actual samples

[0037] ...

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Abstract

A method for detecting aflatoxin B1 based on hybridization chain reaction and DNAzyme signal amplification fluorescence detection, the invention relates to the construction and application of a signal amplification technology fluorescence spectrum sensor. Using the target aflatoxin B1 to trigger a change in the conformation of the aptamer DNA, promote the assembly of the hairpin probe to form a long-chain repetitive DNAzyme unit, and catalyze the enzyme-free AFB1 sensing strategy that cuts the molecular light to generate a fluorescent signal. Relying on this method, the detection limit can reach 0.5 ng / mL, which is far lower than the national standard (10 ng / mL). In addition, we tested aflatoxin B1 in the moldy rice extract and also obtained satisfactory results.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and biosensing detection, and relates to the construction and application of a signal amplification technology fluorescence spectrum sensor. In addition, the present invention also relates to a method for detecting aflatoxin B1 by using the fluorescent spectrum sensor. Background technique [0002] Aflatoxin is a mycotoxin produced by Aspergillus flavus, A. It is classified as the first carcinogen by the International Agency for Research on Cancer (IARC). AFB1 widely exists in many contaminated foods, such as grains, beer, spices and their additives, nuts and beans. Many countries have stipulated the content standard of AFB1 in crop products. The European Union stipulates that the content of aflatoxin B1 in food should not exceed 2 ppb. Therefore, the control detection of aflatoxin B1 content is of great significance to human health diet. [0003] Traditional methods for the detection of a...

Claims

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Application Information

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IPC IPC(8): C12Q1/682C12Q1/6825
CPCC12Q1/682C12Q1/6825C12Q2525/205C12Q2563/107C12Q2565/607
Inventor 明晶晶姜廷福吕志华
Owner OCEAN UNIV OF CHINA
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