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Reagent kit for testing corn zeranol and testing method thereof

A technology of zearalenone and kits, which is applied in measuring devices, color/spectral characteristic measurements, instruments, etc., can solve the problems of expensive imported kits and limited scope of use, and achieve simple structure, high sensitivity, and easy to use convenient effect

Active Publication Date: 2007-05-16
无锡市江原实业技贸有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high price of imported kits, its scope of use is limited to a certain extent.

Method used

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  • Reagent kit for testing corn zeranol and testing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 prepares kit and detects corn sample

[0021] Eu 3+ - Preparation of goat anti-mouse antibody:

[0022] Take 2ml of 10g / L goat anti-mouse antibody dissolved in 50mmol / L PBS pH7.0, change the buffer condition through PD-10 column, the eluent is 50mmol / L NaCl containing 0.155mol / L NaCl 2 CO 3 -NaHCO 3 pH 8.5 buffer. The protein peaks were collected and quantified by UV absorption analysis (1.46A280 -0.74A 260 ), dilute the goat anti-mouse antibody to 2g / L with the eluent above. Take 1ml of diluted goat anti-mouse antibody and add 0.5mg of Eu 3+ -N 2 -[p-isocyanate-benzyl]-diethylenetriaminetetraacetic acid (Eu 3+ -DTTA) vial, 30 ℃ magnetic stirring reaction for 20 hours. The reaction solution was chromatographed on a Sepharose CL-6B column (1×40cm) equilibrated with 80mmol / L Tris-HCl pH7.8 buffer solution, A 280 Monitor the collected protein peaks and dilute the aliquots for use.

[0023] Preparation of solid-phase antigen on coated plate:

[002...

Embodiment 2

[0055] Example 2 Determination of ZEN content in wheat

[0056] Operation method is the same as embodiment 1. The Cps value is 1256385. Calculate the ZEN content in the sample from the standard curve, see Table 1 and Figure 2, the ZEN concentration in the sample in this example is 0.51ng / ml.

Embodiment 3

[0057] Example 3 Determination of ZEN content in corn

[0058] Operation method is the same as embodiment 1. The Cps value is 1905218. Calculate the ZEN content in the sample from the standard curve, see Table 1 and Figure 2, the ZEN concentration in the sample in this example is 0.07ng / ml.

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PUM

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Abstract

This invention relates to one agent case and its test method for corn zeranol, which is based on label immune reaction, wherein, the micro hole board comprises ZEN-BSA added with ZEN antibody; the free ZEN and micro board ZEN-BSA competes for ZEN antibody and the free ZEN antibody is removed with labeled sheep antipest antibody; the labeled immune reaction labeled antibody is removed; after adding strengthen liquid it uses time resolution fluorescence device to test the intensity cps with its concentration reverse to ZEN concentration to determine the ZEN content by comparing standard curve and sample.

Description

technical field [0001] A kit for detecting zearalenone and a detection method thereof belong to the technical field of time-resolved fluorescence immunoassay (TRFIA), and are used for detecting the content of zearalenone (ZEN for short) in grains. Background technique [0002] ZEN is an estrogenic mycotoxin produced by F. graminearum, F. tricinetum, F. oxysporum, F. culmorum, F. moniliforme F.oeniliforme, F.eguiseti, F.avenaceum, F.nivale and other bacteria. Zearalenone mainly exists in corn and corn products, and is also distributed in wheat, barley, sorghum, and rice to a certain extent. It mainly pollutes wheat, barley, oats, millet, sesame, hay, and silage. Zearalenone has strong reproductive toxicity and teratogenic effects, and can cause estrogen poisoning in animals. The main symptoms are vaginal and mammary gland swelling, uterine enlargement, eversion, resulting in infertility or abortion in animals, especially for poultry. Pigs, cattle and sheep have a greater im...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/25
Inventor 黄飚金坚屠蔷陶文沂时瑾王雪
Owner 无锡市江原实业技贸有限公司
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