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Chemiluminescence kit for detecting Fumonisins and preparation method thereof

A technology of fumonisin and chemiluminescence enzyme, which is applied in the field of immunodetection, can solve the problems of complex extraction and purification steps, difficult to popularize and use, restricted sensitivity and the like, and achieves the effects of high sensitivity, improved sensitivity, and simple and convenient operation.

Inactive Publication Date: 2012-05-30
JILIN UNIV
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Instrument analysis methods such as HPLC and capillary electrophoresis have high sensitivity and specificity, but they require complicated extraction and purification steps and expensive instruments, which are not easy to popularize and use
Although the ELISA method is simple, fast, and sensitive, it is also suitable for large-scale screening work, but its sensitivity has certain restrictions.

Method used

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  • Chemiluminescence kit for detecting Fumonisins and preparation method thereof
  • Chemiluminescence kit for detecting Fumonisins and preparation method thereof

Examples

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Embodiment 1

[0030] 1. Antigen preparation

[0031] 1.1 Preparation of immune antigen

[0032] Fumonisin was coupled to hemocyanin by the glutaraldehyde method. The specific operation was as follows: 1.0 mg KLH and 600 μg FB1 were dissolved in 1.0 mL of 0.01 mol / L phosphate buffer solution, and an equal volume of 0.5% glutaraldehyde was added. Aldehyde solution, stirred at room temperature for 2 hours, added 0.25 mL of 1 mol / L glycine, continued to stir for 1 hour, dialyzed and concentrated, redissolved in 0.01 mol / L phosphate buffer solution, and stored at -20°C.

[0033] 1.2 Preparation of coated antigen

[0034] Fumonisin was coupled to ovalbumin by the glutaraldehyde method. The specific operation was as follows: 1.5 mg OVA and 300 μg FB1 were dissolved in 1.5 mL 0.01 mol / L phosphate buffer solution, and an equal volume of 0.5% glutaraldehyde was added. Aldehyde solution, stirred at room temperature for 2 hours, added 0.375 mL of 1 mol / L glycine, continued stirring for 1 hour, dial...

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Abstract

The invention relates to a chemiluminescence enzyme linked immunosorbent assay kit for detecting Fumonisins. The chemiluminescence kit has the characteristics: a luminescent plate is a detachable 96 aperture opaque white luminescent plate adsorbing a conjugate of Fumonisins and ovalbumin (OVA), and a conjugate coating concentration is 0.25mg / L; a standard substance concentration is 0, 0.1, 0.5, 2.5, 12.25, 61.25 mug / L; a ratio of methanol to PBS is 7:93 (v / v); a chemiluminescence substrate liquid is a mixed solution of luminol and superoxol in a ratio of 1:1; a weak solution is a phosphatic buffer (0.01M, pH 7.4) containing 0.5% BSA; a 10 time concentrated washing liquid is 0.1M phosphatic buffer (containing 0.5% tween-20). The chemiluminescence kit has characteristics of high sensitivity, strong singularity and easy and convenient operation, and can be used to detection of Fumonisins in grain of cereal and products thereof.

Description

technical field [0001] The invention discloses a fumonisin chemiluminescent enzyme-linked immunoassay kit, and also provides a preparation method of the kit, which belongs to the field of immunoassay. Background technique [0002] Fumonisins are a class of mycotoxins mainly produced by Fusarium Moniliforme discovered in the late 1980s. So far, fumonisins including fumonisins B1 (FB1 ), fumonisin B2 (FB2) and fumonisin B3 (FB3), etc., among which FB1 is the main component of fumonisin in contaminated corn or Fusarium moniliforme cultures, and is also the cause of fumonisin The main cause of the toxic effect of equine. The relative molecular mass of FB1 is 721.83. Among the numerous mycotoxins, fumonisins are considered to be one of the more serious threats to human and animal health. It is widely found in corn and its products worldwide, and is related to the high incidence of human esophageal cancer. Can cause equine leukomalacia (ELEM), porcine pulmonary edema syndrome (...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543G01N21/76
Inventor 李岩松李小兵柳增善刘国文杨威张燚孔涛唐佳佳李冬娜王哲
Owner JILIN UNIV
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