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Quantitative detection method of fumonisins B1

A technology of fumonisins and FB1-OVA, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of unfavorable routine detection and use, and achieve the effects of easy promotion and application at the grassroots level, strong pertinence, and short time required

Inactive Publication Date: 2012-06-27
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, this method requires strict pretreatment of the test samples, high-performance liquid chromatography and other expensive instruments, and requires professionals to operate, which is not conducive to on-site routine testing.

Method used

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  • Quantitative detection method of fumonisins B1
  • Quantitative detection method of fumonisins B1
  • Quantitative detection method of fumonisins B1

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Experimental program
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Embodiment

[0031] 1. Antigen preparation

[0032] (1) Preparation of fumonisin B1 immune antigen

[0033]Put 1ml of KLH (10mg / ml) into a dialysis bag, place it in 200ml of PBS solution containing 0.2% (V / V) glutaraldehyde (GA) and dialyze at 4°C for 16h, then transfer to PBS and dialyze for 8h to remove unreacted Ga. Add 1 mg of FB1 to the KLH dialyzate and react at 4°C for 16 hours. Add 10 mg Tris, react for 2 hours, block unreacted protein sites, finally dialyze with PBS for 2-3 days, and store at -20°C to obtain the complete FB1-KLH antigen.

[0034] (2) Preparation of Fumonisin B1 Coating Antigen

[0035] Dissolve 2.5 mg ovalbumin (OVA) in 0.1 ml 0.01 M PBS buffer, add 10 μl 50% (V / V) GA, and stir overnight at room temperature. Dialyze against PBS overnight at 4°C to remove excess GA. Take 0.5mg of fumonisin (FB1) and dissolve it in 0.2ml of 25% (V / V) ethanol, add it to the activated OVA dialyzate (about 0.15ml), add 0.1ml of 1M carbonic acid buffer (pH9.5), Stir overnight at 4...

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Abstract

The invention relates to a quantitative detection method of fumonisins B1, comprising the following steps of: 1, preparing conjugates FB1-KLH and FB1-OVA; 2, preparing anti-FB1 monoclonal antibody; 3, preparing colloid gold, labeling the anti-FB1 monoclonal antibody and spraying the labeled monoclonal antibody onto a gold labeled gasket; 4, spotting on a nitrocellulose membrane; 5, assembling to form strips; and 6, drawing a standard curve of FB1 concentration and color value and dragging the color value of a sample to be tested into the standard curve to obtain the FB1 content in the sample to be tested. The invention has advantages of single detection object and strong pertinence, high accuracy, fast detection speed and short required time. The method provided by the invention can be used for detection without trained professionals. The method satisfies the demand of rapid and correct judgment of FB1 content in food storage and sale institution, exit-entry and custom inspection departments, and is convenient for popularization and application at grass-root level.

Description

technical field [0001] The invention relates to a method in the technical field of biological detection, in particular to a method for quantitatively detecting fumonisin B1. Background technique [0002] Fumonisin B1 (Fumonisin B1, FB1) is a secondary metabolite produced by Fusarium moniliforme under certain humidity and temperature conditions, and it is widely distributed all over the world. In 1988, Gelderblom et al first isolated fumonisins from the culture solution of Fusarium moniliforme. Fumonisins can contaminate corn and its products, and have been detected in some grain-based products such as noodles, beer, condiments, and even in asparagus. Fumonisins have been reported to be associated with equine leukomalacia, pulmonary edema syndrome in pigs, liver cancer in rats and other diseases. In addition to the above-mentioned diseases, studies in the Transkei region of South Africa and the Linxian region of China have found that consumption of fumonisin-contaminated co...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 严亚贤王元凯孙建和
Owner SHANGHAI JIAO TONG UNIV
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