Method for detecting fungaltoxin by using quantum dot cation exchange signal amplification technology

A cation exchange and mycotoxin technology, applied in the field of bioengineering, can solve the problems affecting the uniformity and accuracy of the results, the limited detection sensitivity of the enzyme standard method, and the interference of sensitivity and stability, so as to shorten the detection time and improve the detection sensitivity and the effect of improving stability and sensitivity

Inactive Publication Date: 2015-10-21
ZHEJIANG UNIV
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Problems solved by technology

[0003] Routine detection methods for mycotoxins mainly include instrumental analysis and enzyme-linked immunosorbent assay (ELISA), among which instrumental analysis cannot be widely used due to the disadvantages of expensive equipment and complicated operation
The operation process of the ELISA detection method based on antigen-antibody reaction is relatively simple, and it has become a routine method for laboratory diagnosis at this stage, but its sensitivity and stability are easily inte

Method used

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Examples

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Example Embodiment

[0031] Example: Rapid detection of ochratoxin OTA using competitive ELISA

[0032] Taking Aspergillus ochrae OTA monoclonal antibody and quantum dots (carboxyl modified water-soluble quantum dots, composed of CdSe / ZnS, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd., catalog number Q2625) as examples, the explanation is based on quantum dot cation exchange Techniques for rapid detection of ochratoxin OTA:

[0033] (1) Coupling of quantum dots and ochratoxin A monoclonal antibody

[0034] Take the purified anti-ochratoxin A monoclonal antibody (mouse source, purified with saturated ammonium sulfate, the purity is more than 90%, the concentration is 1.5mg / ml, the solvent is 0.01M phosphate buffer PBS, pH=7.4). Take 50μl of carboxy-modified quantum dots with a concentration of 8μM and place them in a 2ml EP tube, then add 135μl of 10mM borate buffer and continue to mix, then add 200μl of the above purified anti-ochratoxin A monoclonal antibody and continue mix...

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Abstract

The invention belongs to the field of bioengineering, and aims to provide a method for detecting fungaltoxin by using a quantum dot cation exchange signal amplification technology. The method includes the following steps: using carboxyl-modified water-soluble cadmium selenide/zinc sulfide quantum dots to mark monoclonal antibodies of fungaltoxin, and enabling carboxyl on the quantum dots and amidogen of the antibodies to form stable fungaltoxin monoclonal antibodies marked by quantum dots under the action of a coupling agent; then detecting fungaltoxin in competitive ELISA mode, and finally analyzing fluorescence signals after reaction by adopting the cation exchange signal amplification technology, so as to realize rapid qualitative detection and quantitative analysis on fungaltoxin. According to the method provided by the invention, the detection time is shortened while the detection sensitivity and stability are improved; the detection of fungaltoxin only needs 45 minutes; the detection sensitivity for the to-be-detected fungaltoxin can reach a picogram level; the quantum dots used in the invention are convenient to synthesize and low in cost; the method has the advantages of being efficient, fast, high in sensitivity, stable in detecting signals, and the like.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method for detecting mycotoxins based on quantum dot cation exchange and signal amplification technology. Background technique [0002] Biotoxins, also known as natural toxins, refer to various chemical substances that are derived from organisms, including animals, plants, and microorganisms, that are toxic to other biological species and cannot replicate by themselves. Human beings' earliest experience with biotoxins stems from frequent food poisoning incidents, and most of the food poisoning causes are biotoxins, and microbial toxins are the most common. Microbial toxins include protein toxoids and mycotoxins. Mycotoxins are small molecular secondary metabolites produced by fungi, which can induce a variety of diseases and are important pollutants in food and feed. The common aflatoxin B1 ( AFB1), ochratoxin A (OTA), and zearalenone (ZEN) all have carcinogenic, tera...

Claims

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Application Information

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IPC IPC(8): G01N33/58
CPCG01N33/588
Inventor 方维焕章先李肖梁谢珲王歆孙孟娇
Owner ZHEJIANG UNIV
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