Electrochemical detection method for AFB1 (Aflatoxin B1) based on strand displacement amplification and surface proximity hybridization reaction and application thereof

A technology of strand displacement amplification and hybridization reaction is applied in the field of electrochemical detection of AFB1 based on strand displacement amplification and surface proximity hybridization reaction, which can solve the problems of difficult operation, low sensitivity of detection method and high detection cost, and achieves simple and easy operation. Batch analysis, the effect of low synthesis cost

Active Publication Date: 2017-05-17
河南省农业科学院农业质量标准与检测技术研究所
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Problems solved by technology

[0004] The present invention is to solve current AFB 1 The detection method has low sensitivity, difficult operation, and high detection cost technical problems.

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  • Electrochemical detection method for AFB1 (Aflatoxin B1) based on strand displacement amplification and surface proximity hybridization reaction and application thereof
  • Electrochemical detection method for AFB1 (Aflatoxin B1) based on strand displacement amplification and surface proximity hybridization reaction and application thereof
  • Electrochemical detection method for AFB1 (Aflatoxin B1) based on strand displacement amplification and surface proximity hybridization reaction and application thereof

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[0026] An AFB Based on Strand Displacement Amplification and Surface Proximity Hybridization Reaction 1 Electrochemical detection method, described detection method will AFB 1 The nucleic acid aptamer is used as the molecular recognition element, and the screen-printed electrode is used as the transducer. After the signal is amplified by the constant temperature strand displacement amplification method, the signal is output through the surface proximity hybridization reaction.

[0027] The AFB 1 The sequence of the nucleic acid aptamer is shown in SEQ ID NO:1.

[0028] AFB Based on Strand Displacement Amplification and Surface Proximity Hybridization Reaction 1 An electrochemical detection method, comprising the following steps:

[0029] (1) Surface modification of magnetic beads: biotin-labeled AFB 1 Nucleic acid aptamer and Block probe stock solutions were pretreated at 95°C for 5min and 0°C for 10min, respectively, and then treated with AFB 1 Dilute 1×BB buffer to 200...

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Abstract

The invention belongs to the technical field of the quality safety detection of agricultural products, and relates to the detection of AFB1 (Aflatoxin B1) in the agricultural products, in particular to an electrochemical detection method for the AFB1 based on a strand displacement amplification and surface proximity hybridization reaction and application thereof. A hybrid complex formed by a biotin-labelled AFB1 aptamer and a Block probe is prefixed on a surface of a magnetic bead; in the presence of a target, the AFB1 and the Block probe are combined with the aptamer in a competitive manner; the replaced Block probe can trigger a strand displacement amplification reaction so as to produce a lot of amplicons; and the amplicons can cooperatively participate in the surface proximity hybridization reaction on a surface of a screen-printed electrode, so as to introduce an electroactive probe to generate a corresponding electrical signal. Compared with specific experiments and conventional ELISA (Enzyme-linked Immuno Sorbent Assay) methods, the electrochemical detection method for the AFB1 disclosed by the invention has the advantages of good specificity, high sensitivity, reliability and accuracy, and is expected to provide a technical selection for large-scale screening of the AFB1 in agricultural products including corn.

Description

technical field [0001] The invention belongs to the technical field of agricultural product quality and safety detection, and relates to aflatoxin B in agricultural products 1 (AFB 1 ) detection, specifically involving an AFB based on strand displacement amplification and surface proximity hybridization reactions 1 Electrochemical detection methods and applications. Background technique [0002] Aflatoxin (AF), as a secondary metabolite of many molds, can contaminate most agricultural products in the field and during storage, such as grains, peanuts, corn, oilseeds and other grains and feeds, and has become an important factor affecting food safety. , a giant killer that endangers the health of humans and animals. The aflatoxins that have been discovered so far have the effects of inducing mutations, suppressing immunity and carcinogenesis, and have been classified as Class I carcinogens by the Cancer Research Institute of the World Health Organization. AFB has been found...

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Application Information

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IPC IPC(8): G01N27/327
CPCG01N27/3275
Inventor 王红旗刘继红张玲李淑芳刘冬梅张军锋贾斌孙江南尚兵
Owner 河南省农业科学院农业质量标准与检测技术研究所
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