Method for detecting ochratoxin A

A technology of ochratoxin and fluorescent signal, which is applied in the field of high-throughput, low-cost detection, and rapid mycotoxin detection. It can solve the problems of expensive instruments, low sensitivity, and complicated sample pretreatment, and achieve stable detection, low cost, and good detection. Selected effect

Inactive Publication Date: 2014-01-22
NANJING NORMAL UNIVERSITY
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Problems solved by technology

These commonly used detection methods play a very important role in the detection of ochratoxin A, but these methods have more or less defects such as: complicated sample pretreatment, expensive instruments, low sensitivity, etc., and with the development of science and technology As well as the improvement of the detection level, the requirements for the limit standard of ochratoxin A in food are also continuously increasing, and the traditional detection methods can no longer meet the current detection needs

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  • Method for detecting ochratoxin A
  • Method for detecting ochratoxin A
  • Method for detecting ochratoxin A

Examples

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Embodiment 1

[0033] The rice, corn and wheat purchased in the market were respectively crushed with a traditional Chinese medicine grinder, passed through a 200-mesh sieve, and 5g of each sample was weighed in a cleaned Erlenmeyer flask with a stopper, and different concentrations and volumes of ocher were added to the sample. Aspergillus toxin A-methanol solution standard, after mixing the sample thoroughly, put it in a ventilated place until the solvent evaporates completely. Add 15mL of extractant (methanol:water=4:6) to it, coat the cork with a layer of water, and cap the cork tightly to prevent leakage. Extract by shaking in a shaker for 2 hours (180r / min), filter with whatman filter paper, take the filtrate and centrifuge for 10min (4000r / p), and take the supernatant, which is the sample to be tested. At the same time do a blank test.

[0034] Detection of ochratoxin A in samples:

[0035] Put the modified microspheres in a centrifuge tube, 5 balls per tube, and then add 10 μL of o...

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Abstract

The invention relates to a method for detecting ochratoxin A. The method utilizes a nucleic acid aptamer sequence fixed on a photonic crystal microsphere and a fluorescence labeled complementary sequence thereof for hybridization; after addition of a sample to be detected, a chip scanner is employed to detect fluorescence signals before and after addition of the sample; and change value of the measured fluorescence signal is used for detection of ochratoxin A in the sample. The method has detection limit of 0.001 ng / mL on ochratoxin A, linear detection range from 0.001 to 1 ng / mL; and only 2 muL of specimen is required.

Description

technical field [0001] The invention relates to a rapid, high-throughput and low-cost detection method for mycotoxins in agricultural products, feed and food. Specifically, the typical ochratoxin A in grains is used as the detection object, and the three-dimensional photonic crystal microspheres are used as the carrier to construct a liquid phase chip detection technology through the combination of nucleic acid aptamer recognition technology and fluorescence analysis technology. Background technique [0002] Ochratoxin is a toxic secondary metabolite produced by toxigenic strains such as Aspergillus and Penicillium. There are many kinds of fungi that produce ochratoxin in nature, mainly Penicillium verrucosum, Aspergillus sochraceus, and Aspergillus carbonarius. Ochratoxins mainly include 7 structural analogs such as A, B, C, and D, among which ochratoxin A (ochratoxin A, OTA) is the most toxic, the most common, and the most widely distributed. OTA has a wide range of food...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/52
CPCG01N33/54366G01N33/54313G01N33/54393
Inventor 李建林孙越徐坤李玮徐杰蒋云坤曹斌郑铁松
Owner NANJING NORMAL UNIVERSITY
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