Label-free visualization detection method and label-free visualization detection kit of aflatoxin B1

An aflatoxin and detection kit technology, which is applied in the direction of chemical reaction of materials and material analysis by observing the influence of chemical indicators, etc., can solve the problems of limited wide application, troublesome storage and high detection cost. , to achieve the effect of being suitable for widespread promotion, economical and low-cost

Inactive Publication Date: 2015-10-07
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Traditional AFB1 detection methods mainly include thin-layer chromatography, high performance liquid chromatography, mass spectrometry, etc. These methods require cumbersome sample pretreatment and expensive instruments, high detection costs, time-consuming and labor-intensive, and require experienced operators Difficult to achieve rapid on-site analysis of large batches of samples
Another method is to use antibodies to detect AFB1, but the preparation process of antibodies is cumbersome, troublesome to store, and involves multiple washing and separation processes, thus limiting their wide application

Method used

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  • Label-free visualization detection method and label-free visualization detection kit of aflatoxin B1

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Experimental program
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Effect test

Embodiment 1

[0028] A label-free visual detection kit for aflatoxin B1 comprises the following components:

[0029] (1) The nucleic acid aptamer of AFB1, the sequence is as follows:

[0030] 5'-GTTGGGCACGTGTTGTCTCTCTGTGTCTCGTGCCCTTCGCTAGGCCCACA-3' (SEQ ID NO. 1);

[0031] (2) Colloidal gold solution;

[0032] (3) 50 mM Tris-HCl solution (pH 7.4);

[0033] (4) 10 mM NaCl solution.

[0034] A label-free visual detection method for aflatoxin B1, comprising the steps of:

[0035] (1) AFB1 aptamer was dissolved in 50 mM Tris-HCl solution (pH 7.4) at a concentration of 20 nM;

[0036] (2) Add colloidal gold solution with a particle size of about 15 nM, mix well, and place at room temperature for 15 minutes;

[0037] (3) Add the sample to be tested, mix well, and react at room temperature for 30 minutes;

[0038] (4) Add 10 mM NaCl solution, mix well, react at room temperature for 10 minutes, observe the color change, the solution remains red, indicating that there is no aflatoxin B1 in the...

Embodiment 2

[0041] Detection of different concentrations of AFB1:

[0042] Prepare AFB1 standard solutions with concentrations of 0.1 ng / mL, 1 ng / mL, 10 ng / mL, 100 ng / mL, and 200 ng / mL, and store at 4 degrees.

[0043] AFB1 solutions of different concentrations were added to the reaction system described in Example 1, and the experimental results were observed after sufficient reaction, such as figure 2 As shown, 0.1 ng / mL of AFB1 can produce a clear blue color change, indicating that its detection limit is 0.1 ng / mL. As the concentration of AFB1 increased, the blue intensity increased and gradually became saturated.

Embodiment 3

[0045] Specificity experiment:

[0046] Prepare standard solutions of different interfering substances with a concentration of 100 ng / mL, namely aflatoxin M1, aflatoxin G1, ochratoxin A, zearalenone, patulin, citrinin and versicolor.

[0047] Add 100 ng / mL different interfering substance standard solutions and 1 ng / mL AFB1 standard solution to the reaction system described in Example 1 respectively, and observe the color change after sufficient reaction, as shown in image 3 As shown, 100 ng / mL of aflatoxin M1, aflatoxin G1, ochratoxin A, zearalenone, patulin, citrinin and versicolor remained red and had no effect on the detection. Only when AFB1 was added, the blue color change occurred, which proved that the method has good specificity for the detection of AFB1.

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Abstract

The present invention discloses a label-free visualization detection method and a label-free visualization detection kit of aflatoxin B1. According to the present invention, the nucleic acid aptamer of aflatoxin B1 is adopted as the molecule recognition element, colloidal gold is adopted as the signal report molecule, the nucleic acid aptamer is added to the gold colloid solution, and the aflatoxin B1 interacts with the nucleic acid aptamer in the presence of the aflatoxin B1 so as to lose the protection effect of the nucleic acid aptamer on the aflatoxin B1; after NaCl is added, the red color of the detection system is changed into the blue and purple color; and if no aflatoxin B1 exists in the system, the red color of the solution is remained. According to the method of the present invention, the detection result is directly visible without any detection equipment, the operation is simple without antibodies, any label modification and any washing separation processes, and the method is suitable for on-site rapid detection, further has characteristics of low cost, economy, low price, fast response, and is suitable for wide promotion.

Description

technical field [0001] The invention belongs to the field of food safety detection and relates to a label-free visual detection and detection kit of aflatoxin B1. Background technique [0002] Aflatoxin B1 (Aflatoxin B1, AFB1) is a secondary metabolite of fungi and belongs to mycotoxins. It is mainly produced by Aspergillus flavus, Aspergillus parasitica and Aspergillus tererus. It is commonly found in moldy grain and oil products and seriously endangers food safety and human body. healthy. AFB1 has strong toxicity and carcinogenicity, and is currently the strongest carcinogenic mutagen among chemical carcinogens, and its toxicity is 10 times stronger than potassium cyanide. AFB1 is listed as a class I carcinogen by the International Agency for Oncology Research. Therefore, the rapid detection of AFB1 is of great significance. Traditional AFB1 detection methods mainly include thin-layer chromatography, high performance liquid chromatography, mass spectrometry, etc. These ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 陈俊华周顺桂
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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