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A kind of detection method for ochratoxin a

A technology of ochratoxin and detection method, which is applied in the field of detection of ochratoxin A, can solve the problems of low precision and achieve the effects of saving cost and improving detection sensitivity

Active Publication Date: 2017-11-14
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the technical defect of prior art, provides a kind of detection method for ochratoxin A, to solve the low accuracy of ELISA detection method of ochratoxin A of prior art

Method used

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  • A kind of detection method for ochratoxin a
  • A kind of detection method for ochratoxin a
  • A kind of detection method for ochratoxin a

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 (a kit was prepared based on the novel fluorescent ELISA detection method of ochratoxin A of the present invention, and it was used to detect the residual amount of ochratoxin A in corn and corn products)

[0054] Preparation and detection method of a new fluorescent ELISA kit for detecting ochratoxin A, including 96-well black fluorescent microtiter plate coated with anti-ochratoxin A monoclonal antibody, ochratoxin A standard substance, and catalase-labeled ochratoxin A working solution, substrate solution A hydrogen peroxide, fluorescent substrate solution B mercaptopropionic acid modified cadmium telluride quantum dots and concentrated washing solution.

[0055] The following specifically describes the preparation of the ELISA kit for detecting ochratoxin A in the present invention,

[0056] The 96-well black fluorescent microtiter plate was purchased from Corning Company of the United States; the anti-ochratoxin A monoclonal antibody was purchased from Wu...

Embodiment 2

[0089] Embodiment 2 (using horseradish peroxidase as an antibody labeling enzyme and using TMB as a chromogenic substrate for the direct competition ELISA detection method of ochratoxin A)

[0090] When the traditional ELISA kit is used to detect the residual amount of ochratoxin A in corn and corn products, it is implemented through the following steps: sample pretreatment, detection with the kit of the present invention, and analysis of the results.

[0091] (1) Sample pretreatment

[0092] Take the pulverized sample and pass it through a 20-mesh sieve, and mix thoroughly; weigh 5g sample, add 12.5mL extract solution (methanol:water=7:3), shake vigorously for 30min; centrifuge at 5000rpm for 10min, filter with filter paper; take 1mL filtrate for Dilute with 1mL PBS and set aside.

[0093] (2) Use a traditional ELISA kit to detect the residual amount of ochratoxin A in the above samples

[0094] Take the ELISA plate coated with anti-ochratoxin A monoclonal antibody, add 50 ...

Embodiment 3

[0100] The invention relates to a detection method for ochratoxin A, which belongs to the direct competition enzyme-linked immunosorbent method, and the method is for the detection of ochratoxin A antigen, and the enzyme used for labeling the antigen in the method is catalase C100.

[0101] On the basis of the above technical solutions, the following conditions are met:

[0102] The substrate in the method includes hydrogen peroxide and mercaptopropionic acid modified cadmium telluride quantum dots.

[0103] The specific detection method includes the following steps:

[0104] 1) Coating anti-ochratoxin A monoclonal antibody, and then adding the sample to be tested;

[0105] 2) Then add catalase C100-labeled ochratoxin A, mix and react in a dark environment at 35°C for 40 minutes, and wash;

[0106] 3) Then add hydrogen peroxide solution with a concentration of 8 μmol / L to mix, and react for 20 minutes at 35°C in a dark environment;

[0107] 4) Then add mercaptopropionic aci...

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Abstract

The invention provides a detection method for ochratoxin A. The method comprises the steps: based on a direct competition ELISA technique, firstly coating a monoclonal antibody, adding a to-be-tested sample and catalase C100 labeled ochratoxin A, combining the ochratoxin A in the sample with a monoclonal antibody competing with the catalase labeled ochratoxin A and fixed on an ELISA plate, catalyzing decomposition of hydrogen peroxide by the catalase, reducing fluorescence quenching of mercaptopropionic acid modified cadmium telluride quantum dots, and determining the content of ochratoxin A in the sample according to the fluorescence intensity. The new catalase is innovatively introduced, the reaction accuracy is improved while the costs are reduced; and at the same time, the more sensitive novel fluorescent substrate mercaptopropionic acid modified cadmium telluride quantum dots are used, and the luminous sensitivity is greatly improved compared with that of a traditional TMB substrate.

Description

technical field [0001] The invention relates to the technical field of antigen detection, and further relates to an ELISA-based antigen detection technology, in particular to a detection method for ochratoxin A. Background technique [0002] Ochratoxins are secondary metabolites produced by Aspergillus chraceors, Pmicilium vermculosutm, Penicilium viridicatum, and several other fungi of the genus Penicillium. Ochratoxin is a series of isocoumarin derivatives, including A, B, C and other 7 compounds with similar structures. Among them, ochratoxin A is the most toxic, can damage the liver and kidney of animals, and has teratogenic, carcinogenic and mutagenic effects, and is considered to be related to the occurrence of human Balkan nephropathy. The International Agency for Research on Cancer (IARC) has classified ochratoxin A as a Class 2B carcinogen. Ochratoxin A often contaminates crops such as wheat, corn, cereals, coffee and grapes. Therefore, establishing a sensitive a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543
CPCG01N33/54306
Inventor 熊勇华江湖陈锐梁毅周耀峰黄小林许恒毅
Owner NANCHANG UNIV