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Chemiluminescent immunoassay method for rapidly detecting zearalenone toxin

A technology of zearalenone and immunochemistry, which is applied in the direction of chemiluminescence/bioluminescence, analysis by making materials undergo chemical reactions, and measurement devices, can solve the problems of unfavorable routine detection and use, and achieve high accuracy and sensitivity High, targeted effects

Inactive Publication Date: 2012-01-11
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

However, these methods require strict pretreatment of test samples, expensive instruments such as high performance liquid chromatography, and professional operators, which are not conducive to on-site routine testing.

Method used

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  • Chemiluminescent immunoassay method for rapidly detecting zearalenone toxin
  • Chemiluminescent immunoassay method for rapidly detecting zearalenone toxin
  • Chemiluminescent immunoassay method for rapidly detecting zearalenone toxin

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Embodiment

[0028] 1. Antigen preparation, preparation of zearalenone-coated antigen (ZEN-OVA)

[0029] Take 0.33ml (3mg / ml) ZEN, mix it in 1.2ml pyridine, add 2mg O-carboxymethyl hydroxylamine, and stir at room temperature for 24h. After vacuum drying, 4 ml of distilled water was added and dissolved to adjust the pH to 8.0. The unreacted ZEN was removed by extraction with benzene (3 ml benzene, extracted 3 times in total), the benzene phase was removed and the water phase was retained. Adjust the pH of the aqueous phase to 3.0, extract with ethyl acetate (10ml ethyl acetate, a total of 4 extractions), extract the ester phase, and discard the aqueous phase. The ester phase was filtered with anhydrous sodium sulfate and dried. The dried crystals were dissolved in 0.5 ml of dioxane treated with basic alumina. Weigh 10mg OVA and dissolve it in 0.7ml 0.05mol / L (pH7.2) PBS solution, and mix the two solutions slowly at 4°C. 1 mg of NHS and 2 mg of DCC were dissolved in 0.2 ml of dioxane, ...

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Abstract

A chemiluminescence immunoassay method for detecting zearalenone toxin in the technical field of biological detection engineering comprises the following steps of: linking ZEN semiantigen with OVA to obtain ZEN-OVA; extracting a sample to be measured to obtain an extract; coating ZEN-OVA on a 96-well white plate at the optimum coating concentration, followed by sealing, respectively adding a known solution of the ZEN pure product and anti-zearalenone monoclonal antibody and a mixed solution of the extract of the sample to be measured and anti-zearalenone monoclonal antibody at different concentrations, followed by incubation; adding HRP-labeled secondary antibody after incubation, and adding a luminescence substrate for luminescent determination to obtain a result; making a standard curve; and acquiring the ZEN concentration in the extract of the sample to be measured through the standard curve. The method provided by the invention can be used to rapidly detect the amount of ZEN in the sample to be measured; in the meanwhile, the method has strong pertinency of detection object and results in high accuracy.

Description

technical field [0001] The invention relates to a method in the technical field of biological detection engineering, in particular to an immunochemiluminescent method for rapidly detecting zearalenone toxin. Background technique [0002] Zearalenone (ZEN) is a secondary metabolite produced by Fusarium fungus under certain humidity and temperature conditions. Zearalenone toxin can exist in corn, wheat, barley, sorghum, rye and other grains, and can also exist in animal tissues that eat ZEN-containing feed, including milk and eggs. Zearalenone toxin is associated with spontaneous breast cancer, oviduct and uterine edema, hyperplasia, sperm cell aberration, apoptosis and other diseases. Zearalenone toxin has the characteristics of widespread distribution, long residual time, difficult to handle, and enhanced toxicity with other toxins. After joining the WTO, the international trade volume of agricultural products and related foods has increased day by day, and the requiremen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/76
Inventor 孙建和王元凯严亚贤
Owner SHANGHAI JIAO TONG UNIV
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