Method for simultaneous detection of zearalenone and fumonisins

A technology of zearalenone and fumonisins, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve the problems of not being able to detect two toxins at the same time, affecting the detection of mycotoxins, uneconomical and environmental protection, etc. The effect of grassroots promotion and application, fast detection speed and short time required

Inactive Publication Date: 2011-12-14
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Although there are currently detection kits for zearalenone and fumonisin B1, each detection kit can only detect one toxin, and cannot detect two toxins at the same time. The operation is troublesome, uneconomical and environmentally friendly. , has seriously affected the detection of

Method used

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  • Method for simultaneous detection of zearalenone and fumonisins
  • Method for simultaneous detection of zearalenone and fumonisins
  • Method for simultaneous detection of zearalenone and fumonisins

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Embodiment

[0031] , Antigen preparation

[0032] (1) Preparation of zearalenone immune antigen

[0033] Take 0.33ml (3mg / ml) ZEN, mix it in 1.2ml pyridine, add 2mg O-carboxymethylhydroxylamine, stir and react at room temperature for 24h; after drying the solution in vacuum, add 4ml distilled water and dissolve it, 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 0.5ml of dioxane treated with basic alumina; 20mg of BSA was weighed and dissolved in 0.7ml of 0.05mol / L (pH7.2) PBS; after that, the two solutions were dissolved in 4 Mix slowly at ℃ to obtain solution a; disso...

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Abstract

A method for simultaneously detecting zearalenone and fumonisins in the technical field of biological detection engineering, comprising the following steps: 1. Prepare conjugates ZEN-BSA and ZEN-OVA; FB1-KLH and FB1-OVA; 2. Prepare anti-ZEN monoclonal antibody and anti-FB1 monoclonal antibody respectively; 3. Prepare colloidal gold, respectively label anti-ZEN monoclonal antibody and anti-FB1 monoclonal antibody; spray the labeled monoclonal antibody onto the gold label pad ; 4. Spotting on the nitrocellulose membrane; 5. Assemble into a test strip; 6. Drop the methanol extraction supernatant of the sample to be tested into the sample groove of the test strip to obtain the points on the nitrocellulose membrane. The color development result of the sample band, the identification, and the result are obtained. The detection object of the method of the present invention is highly targeted and has high accuracy; the detection speed is fast and the required time is short, the loss of manpower and material resources caused by using two detection methods to detect two toxins is reduced, and it is convenient for promotion and application at the grassroots level.

Description

technical field [0001] The invention relates to a method in the technical field of biological detection engineering, in particular to a method using a nano colloidal gold immune layer [0002] A method for the simultaneous detection of zearalenone and fumonisins by analytical technology. Background technique [0003] Zearalenone (ZEN) is a secondary metabolite produced by some strains of Fusarium under certain humidity and temperature conditions. Zearalenone can be detected in grains such as wheat, barley, corn, rye, sorghum, and can also be detected in some animal tissues or products (such as milk, eggs, etc.). Zearalenone toxin can cause a variety of diseases in humans and animals, mainly carcinogenic and cancer-promoting toxicity. Reproductive system diseases have an important relationship. Zearalenone toxin has the characteristics of widespread distribution, long residual time, difficult to handle, and enhanced toxicity with other toxins. [0004] Fumonisins are seco...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/558
Inventor 严亚贤王元凯陈志飞李树清孙建和
Owner SHANGHAI JIAO TONG UNIV
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