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Production and use of test paper for fast detecting deoxynivalenol in cereal

A technology of deoxynivalenol and test strips, which is applied in the field of preparation of rapid detection test strips, can solve problems such as increased production costs and complex production processes, and achieve high labeling rate, high sensitivity, and broad market prospects Effect

Inactive Publication Date: 2009-07-15
NANCHANG BOHENG BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection antigen of this patent is DON-BSA, in which BSA is not cationic BSA (MBSA), and the NC membrane needs to be sealed during the preparation process, the production process is relatively complicated, which increases the production cost
Deng Shunzhou and others in our laboratory published a research paper on using MBSA-coupled DON as a detection antigen and applying it to ELISA detection (Food Science, 2007, Vol.28, No.02, P149-152; Food Science and Technology, 2006, No.8 , P222-224), but there is no report that MBSA-coupled detection antigen is used for rapid detection of DON

Method used

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  • Production and use of test paper for fast detecting deoxynivalenol in cereal
  • Production and use of test paper for fast detecting deoxynivalenol in cereal
  • Production and use of test paper for fast detecting deoxynivalenol in cereal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Preparation of DON Rapid Detection Trial Strip

[0038] 1. Preparation of DON monoclonal antibody-colloidal gold marker

[0039] (1) Preparation of colloidal gold

[0040] Dilute 1% chloroauric acid to 0.01% with ultrapure water, put it in an oil bath with magnetic stirring and heat to boil, add 10mL 1% trisodium citrate for every 500mL of 0.01% chloroauric acid, continue to boil until the liquid turns red. Heating was stopped, and the evaporated water was replenished after cooling to room temperature to obtain a colloidal gold solution.

[0041] (2) Preparation of DON monoclonal antibody-colloidal gold marker

[0042] Under magnetic stirring, add K to the colloidal gold solution 2 CO 3The pH of the solution was adjusted to 7.5. Add 20 μg / mL antibody colloidal gold to the antibody DON monoclonal antibody, and continue stirring for 15 minutes. Add 10% BSA to a final concentration of 1.0%, and continue stirring for 15 min. Add 5% PEG-20000 to a final co...

Embodiment 2

[0049] Example 2. Detection of DON in samples

[0050] Detection of DON in Rice, Wheat and Corn

[0051] (1) Sample pretreatment

[0052] Weigh 5 g of pulverized samples to be tested (such as wheat, corn, and rice) into a conical flask, add 50 mL of distilled water for extraction, shake for 5 min, quickly filter with qualitative filter paper, and mix 160 μL of filtrate with 40 μL of buffer for detection.

[0053] (2) Analysis of results

[0054] Take 50 μL and slowly drop it on the sample pad of the test strip, observe the color development of the test strip after 10 minutes, and determine the content of DON in the sample according to the detection principle of the test strip. If both the detection line (T line) and the quality control line (C line) are red, the sample is negative (less than 1 μg / mL); if only the T line is red, the sample is positive (greater than 1 μg / mL); if only the T line Color development or no color development on T line and C line indicates that the...

Embodiment 3

[0055] Example 3. (Application example)

[0056] 1. Specificity experiment

[0057] Carry out the experiment according to the method described in Example 2, dilute the concentration of Aspergillus flavus B1, zearalenone, ochratoxin, T-2 toxin to 50ng / mL, detect with this patent test strip, two red lines appear , the result is negative, that is, there is no cross-reaction between the DON test strip and Aspergillus flavus B1, zearalenone, ochratoxin, T-2 toxin, etc.

[0058] 2. Sensitivity experiment

[0059] Using the method described in Implementation 2, use test strips to detect 0, 6.25, 12.5, 25, 50, 100, 200 and 400ng / mL DON standard substances respectively, repeat 10 times, wherein 0, 6.25, 12.5 and 25ng / mL If two red lines appear, it is negative; if only one red line appears at 50, 100, 200 and 400ng / mL, it is positive. Therefore, the sensitivity of this test strip is 50ng / mL.

[0060] 3. Uniformity experiment

[0061] Using the method described in Implementation 2,...

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Abstract

The invention provides a preparation method of test paper scrip capable of quickly detecting deoxynivalenol test paper in grain and use thereof, comprising (1) DON monoclonal antibody-colloidal gold label; (2) coating of conjugate releasing mat; (3) coating of cellulose nitrate film; (4) production of test paper scrip. The preparation method has features of specificity, high sensitivity; visual detection result; simple operation and high speed, no need of professional operation, obtaining the detection result in 10min; easy large-scale popularization and application, widely application on foodstuff purchasing scene detection, food safety detection, customs quarantine control or the like, with wide market prospect, bigger economic, social benefits.

Description

technical field [0001] The invention relates to a rapid detection method of mycotoxins, in particular to the preparation and application of a rapid detection test strip for deoxynivalenol (DON) in grains. Background technique [0002] Deoxynivalenol (DON), also known as vomi toxin, is mainly a toxic metabolite-trichothecenes produced by certain species of Fusarium fungi with similar chemical structure and biological activity one of the compounds. DON is highly toxic and mainly pollutes cereal crops such as wheat, barley, and corn. People and animals can have a wide range of toxic effects after eating food or feed contaminated with this toxin, often causing vomiting, headache, fever and nausea. In recent years, studies have found that DON may be related to human esophageal cancer and IgA nephropathy, posing a threat to human and animal health. At present, various countries have established strict limit standards for the content of DON in ancient objects and feedstuffs. The...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/543G01N33/532
Inventor 许杨黄志兵何庆华刘仁荣许夏
Owner NANCHANG BOHENG BIOLOGICAL PROD
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