Method for detecting fumonisin by colloidal gold immunochromatographic test

A technology of fumonisins and immunochromatography, applied in measuring devices, analytical materials, 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, high accuracy, and a single detection object

Inactive Publication Date: 2010-03-03
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

In addition, this method requires strict pretreatment of the test samples, high-performance liquid chromatography and othe

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  • Method for detecting fumonisin by colloidal gold immunochromatographic test
  • Method for detecting fumonisin by colloidal gold immunochromatographic test

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[0026] 1. Antigen preparation

[0027] ① Preparation of immune antigen: put 1ml KLH (10mg / ml) into a dialysis bag, put it in 200ml PBS solution containing 0.2% (V / V) (glutaraldehyde (GA)) and dialyze at 4°C for 16h, then transfer to PBS Dialyze for 8h to remove unreacted GA. Add 1 mg FB to KLH dialyzate 1 , 4°C for 16h. Add 10mg Tris and react for 2h to block unreacted protein sites. Finally, it was dialyzed with PBS for 2-3 days and stored at -20°C.

[0028] ② Preparation of coated antigen: Dissolve 2.5 mg of ovalbumin (OVA) in 0.1 ml of 0.01 MPB buffer, add 10 ul of 50% (V / V) GA, and stir overnight at room temperature. Dialyze against PBS overnight at 4°C to remove excess GA. 0.5mg fumonisins (FB 1 ) was dissolved in 0.2ml of 25% (V / V) ethanol, added to the activated OVA dialyzate (about 0.15ml), added 0.1ml of 1M carbonic acid buffer (pH9.5), and stirred overnight at 4°C. Add 0.05ml of 1M lysine (pH7), and react at 4°C for 3h. Finally, dialyze with PBS for 72 hours,...

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Abstract

The invention discloses a method for detecting fumonisin by a colloidal gold immunochromatographic test in the technical field of biological detection engineering. The method comprises the following steps: conjugating fumonisin hapten with KLH and OVA respectively; preparing a monoclonal antibody of the fumonisin from the obtained fumonisin-KLH conjugate by a conventional method; preparing 40nm colloidal gold by a trisodium citrate reducing method, and labeling the monoclonal antibody of the fumonisin subject to dialysis desalination treatment with the colloidal gold; spraying the obtained monoclonal antibody protein onto a gold-labeled pad, spraying the fumonisin-OVA conjugate onto a T line, spraying a rabbit antimouse monoclonal antibody onto a C line to form a reagent paper tape, and drying; and extracting a sample to be tested with methanol, centrifuging and taking supernatant, adding the supernatant to a sample groove of the reagent paper tape dropwise to acquire development of the T line and the C line, and identifying to obtain results. The method has advantages of high detection specificity, high accuracy, high detection speed and detection time as short as only 5-10min.

Description

technical field [0001] The invention relates to a method in the technical field of biological detection engineering, in particular to a method for detecting fumonisins by colloidal gold immunochromatography test. Background technique [0002] Fumonisins are secondary metabolites produced by Fusarium moniliforme under certain humidity and temperature conditions, and are 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-...

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Application Information

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