Preparation and detection method for ELISA kit detecting Fumonisins

A fumonisin and detection method technology, applied in the field of detection, can solve problems such as backward development, and achieve the effects of simple operation and sensitive detection

Inactive Publication Date: 2014-05-14
JIANGSU WISE SCI & TECH DEV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, most of the immunological detection kits for fumonisins are imported,

Method used

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  • Preparation and detection method for ELISA kit detecting Fumonisins

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Embodiment Construction

[0032] An ELISA kit for detecting fumonisins, which includes an enzyme-labeled plate, fumonisins standard, fumonisin antibody working solution, fumonisin enzyme-labeled secondary antibody working solution, substrate solution A, substrate solution B, and stop solution And concentrated washing liquid.

[0033] The following specifically describes the preparation of the ELISA kit for detecting fumonisins in the present invention.

[0034] Preparation of Fumonisin Protein Conjugate:

[0035] The fumonisin hapten and the carrier protein BSA were mixed in 0.05 mol / L pH 9.6 carbonate buffer (CBS) at a binding ratio of 12:1, then added to carbodiimide, stirred for 1-2h, set Reaction at room temperature for 24 hours, and finally dialysis with 0.2 mol / L pH 7.6 PBS for two days to remove the unreacted hapten, and the fumonisin protein conjugate can be obtained.

[0036] Preparation of fumonisin antibody:

[0037] Three-month-old healthy white rabbits were selected for three immunizations. The do...

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Abstract

The invention relates to a preparation and detection method for an ELISA kit detecting Fumonisins. The ELISA kit has the characteristics of sensitive, accurate and fast detection, simple operation and strong specificity, and is suitable for detection of a large number of samples. The kit includes: a Fumonisins antigen coated enzyme label plate, a Fumonisins standard substance, a Fumonisins antibody working solution, a Fumonisins enzyme labeled secondary antibody working solution, a substrate solution A, a substrate solution B, a terminating solution, a concentrated sample dilution solution and a concentrated washing solution. The principle of the Fumonisins detection kit is solid-phase indirect competitive enzyme-linked immunosorbent assay reaction. The extracted sample, the enzyme labeled secondary antibody working solution, and the antibody working solution are added into corresponding enzyme labeled holes, after incubation for a period of time, the substrate solution A and the substrate solution B are added into a washing plate, and under the action of enzyme, the holes can present a blue color. Then the terminating solution is added, and the color changes to yellow from blue. The color depth and the content of Fumonisins in the standard substance or the sample are in an inverse proportion relationship. This method can be directly used for detecting Fumonisins in maize.

Description

Technical field [0001] The invention belongs to the technical field of detection, and specifically relates to an ELISA kit and a detection method for quantitatively detecting the residual amount of fumonisins in nuts, grains and grain products. Background technique [0002] Fumonisins are a group of mainly caused by Fusarium moniliforme ( Fusarium moniliforme ) Mycotoxins produced by reproduction under certain temperature and humidity conditions. In 1988, Gelderblom et al. isolated fumonisin B1 from corn for the first time. Fumonisins are widely distributed in nature and have great harmfulness, and gradually attracted the attention of scholars at home and abroad. At present, the structural analogues of fumonisins that have been found are mainly divided into four categories: A, B, C, and P, including FA1, FA2, FB1, FB2, FB3, FB4, FC1, FC2, FC3, FC4, FP1, etc. [0003] Fumonisins (B1, B2, and B3) are mycotoxins produced by Fusarium (mold) Candida. Fumonisins have been found to con...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/56911G01N2333/40
Inventor 杜道林洪霞
Owner JIANGSU WISE SCI & TECH DEV
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