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Method for detecting aflatoxin B1 by enzyme-labeled aptamer

A kind of technology of aflatoxin and nucleic acid aptamer, applied in the direction of measuring device, color/spectral characteristic measurement, instrument, etc., can solve the problems of high preparation cost, sensitive to environmental factors, unstable activity, etc., and achieve rapid analysis and high sensitivity , the effect of great application prospects

Active Publication Date: 2017-09-22
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

However, in enzyme-linked immunoassays, antibodies have some limitations, such as immune antibodies are sensitive to environmental factors, their activity is unstable, and their preparation costs are high, and the reproducibility between experiments of different batches of immune antibody products is low.

Method used

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  • Method for detecting aflatoxin B1 by enzyme-labeled aptamer
  • Method for detecting aflatoxin B1 by enzyme-labeled aptamer
  • Method for detecting aflatoxin B1 by enzyme-labeled aptamer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1, prepare the nucleic acid aptamer of HRP mark

[0067] 1. Prepare a biotin-labeled nucleic acid aptamer (artificially synthesized by Shanghai Bioengineering Co., Ltd.).

[0068] The nucleotide sequence of the nucleic acid aptamer is shown in Sequence 1 of the Sequence Listing, and the 5' end of the nucleic acid aptamer is linked to biotin by using triethylene glycol (TEG) as a connecting arm.

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Abstract

The invention discloses a method for detecting aflatoxin B1 by enzyme-labeled aptamer. The method sequentially includes the steps: (1) taking a coated microporous plate, adding a sample to be detected and a specific probe for co-incubation and then washing the microporous plate; (2) detecting the aflatoxin B1 by detecting corresponding signals of the specific probe. The coated microporous plate is a microporous plate coated with aflatoxin B1-BSA conjugates, the specific probe is a compound of a first element and a second element, the first element is a substance obtained by connecting biotin to the tail end of the aptamer specifically combined with the aflatoxin B1 by the aid of triethylene glycol serving as a connecting arm, and the second element is horseradish peroxidase-labeled streptavidin. The enzyme-linked aptamer analysis method has the advantages of simplicity, rapidness and high sensitivity, a plurality of samples can be rapidly analyzed, a detection kit can be made, and the method has a great application prospect in AFB1 analysis detection.

Description

technical field [0001] The invention relates to a method for detecting aflatoxin B1 by using an enzyme-labeled nucleic acid aptamer. Background technique [0002] Aflatoxin is a kind of mycotoxin, mainly refers to the secondary metabolite produced by Aspergillus flavus and Aspergillus parasiticus. Aflatoxins are present in a variety of contaminated foods such as corn, cereals, alcohol, nuts, seasonings and soy products. Among them, aflatoxin B1 (Aflatoxin B1, AFB1) is the most toxic aflatoxin, and is recognized as a major carcinogen by the International Agency for Research on Cancer. Considering the threat of AFB1 to human health and food safety, and its widespread contamination, sensitive detection of AFB1 is necessary. [0003] Currently, chromatographic analysis methods such as high-performance liquid chromatography, liquid chromatography-tandem mass spectrometry, and thin-layer chromatography have been widely used in the quantitative analysis of AFB1. Chromatographic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N21/31
CPCG01N21/31G01N33/535
Inventor 赵强孙琳琳
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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