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Ochratoxin A hapten, ochratoxin A artificial antigen, preparation method of ochratoxin A hapten, kit, and detection method of ochratoxin A

An ochratoxin and artificial antigen technology, applied in the detection of ochratoxin A and the field of ochratoxin A hapten, can solve the problems of low detection limit of thin layer chromatography and expensive equipment, etc.

Active Publication Date: 2018-11-16
CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The detection limit of thin-layer chromatography is too low, and the high-performance liquid chromatography-mass spectrometry (HPL-CMS) equipment is relatively expensive, and not all testing institutions have

Method used

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  • Ochratoxin A hapten, ochratoxin A artificial antigen, preparation method of ochratoxin A hapten, kit, and detection method of ochratoxin A
  • Ochratoxin A hapten, ochratoxin A artificial antigen, preparation method of ochratoxin A hapten, kit, and detection method of ochratoxin A
  • Ochratoxin A hapten, ochratoxin A artificial antigen, preparation method of ochratoxin A hapten, kit, and detection method of ochratoxin A

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preparation example Construction

[0028] Preferably, the preparation method of the ochratoxin A artificial antigen described in the embodiments of the present invention comprises:

[0029] pretreatment of the ochratoxin A hapten by an activated ester method; and

[0030] The pretreated ochratoxin A hapten is coupled to the first carrier protein.

[0031] Pretreatment of the ochratoxin A hapten by the activated ester method is to esterify the carboxyl group of the ochratoxin A hapten, further preferably, the ochratoxin A hapten and N-hydroxysuccinimide (NHS) and N,N-dicyclohexylcarbodiimide (DCC) are dissolved in dimethylformamide (DMF) for reaction, and more preferably, 20-40 mg of the ochratoxin A hapten is mixed with 10-20ml of N-hydroxysuccinimide (NHS) and N,N-dicyclohexylcarbodiimide (DCC) were reacted. The above reaction process is carried out under the condition of avoiding light at room temperature and continuously stirring for 10-12 hours. Preferably, the above reaction process is continuously stir...

Embodiment 1

[0052] Preparation of Ochratoxin A Hapten

[0053] Weigh 30mg of ochratoxin A and 15ml p-benzoyl chloride and dissolve in 3ml dimethylformamide (DMF), add triethanolamine at 0°C, react for 1 hour, and quench the reaction with sodium bicarbonate solution It is extracted, dried and concentrated to obtain the ochratoxin A hapten.

Embodiment 2

[0055] Preparation of Artificial Antigen of Ochratoxin A

[0056] Weigh 100 μL of the above ochratoxin A hapten, 10 mg NHS and 20 mg DCC, dissolve in 500 μL dimethylformamide (DMF), and stir at room temperature in the dark for 12 h; Thyroglobulin was added to the 10% N,N-dimethylformamide solution while stirring. After 1 hour, transfer to 0.01M PBS at 4°C for dialysis with magnetic stirring for 2 days, during which the dialysate was changed every 6 hours. When there is no small molecule absorption peak in the dialysate by ultraviolet scanning, it can be centrifuged to obtain the supernatant, which is the ochratoxin A artificial immunogen, and the product is divided into centrifuge tubes and stored at -20°C.

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Abstract

The invention provides an ochratoxin A hapten, an ochratoxin A artificial antigen, a preparation method of the ochratoxin A hapten, a kit, and a detection method of ochratoxin A. The chemical structural formula (I) of the ochratoxin A hapten is shown in the description.

Description

technical field [0001] The invention relates to an enzyme-linked immunoassay technology, in particular to an ochratoxin A hapten, an artificial antigen, a preparation method thereof, a kit and a detection method of the ochratoxin A. Background technique [0002] Ochratoxin A (OchratoxinA, OTA) is produced by two types of molds, Aspergillus and Penicillium, and often contaminates food and feed. Ochratoxin A can cause acute and chronic poisoning in animals, is a strong carcinogen, can cause irreversible lethal poisoning to the kidney, cause kidney atrophy, and can also cause fetal malformation, abortion or even death, seriously affecting the function and growth of animals. Potential hazards to humans are of concern. For this reason, the International Agency for Research on Cancer has positioned it as a Group IIB carcinogen, and strengthening the detection of ochratoxin A is an important method to prevent food and feed contaminated by ochratoxin A from directly or indirectly e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/531G01N33/543
CPCG01N33/53G01N33/531G01N33/54306
Inventor 骆鹏杰陈霞周爽赵云峰李敬光张霁月
Owner CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT