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Test paper for testing afatoxin by nontoxic system immunochromatographic techniqul and preparing process thereof

A technology for immunochromatographic detection and aflatoxin, which is applied in measurement devices, analysis materials, and material analysis by optical means, etc., can solve the problems of health hazards, high price, and dependence on imports for production and operators, and avoid two problems. The effect of secondary pollution, easy operation and high sensitivity

Inactive Publication Date: 2005-10-26
ZHONGDE DADI BIOENG JIANGXI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned methods all have the following disadvantages: the operation procedure is complicated and can only be carried out in the laboratory; professional personnel are required to operate, use special instruments and equipment, the detection time is long, and the required cost is high, and the cost is 200-1000 yuan / per month samples
[0006] It is well known that aflatoxins are highly toxic, among which aflatoxin B 1 It is the most toxic natural substance known, which is 10 times more toxic than potassium cyanide. It is very likely that there will be hidden dangers of secondary pollution if aflatoxin is used as a material for production or aflatoxin as a reference substance for detection. The health of production and operating personnel is extremely harmful
Furthermore, the high-concentration toxin standard products required by our country are currently all dependent on imports, which are expensive, making the cost of detection reagents high
What is even more troublesome is that after the "9.11" terrorist attack, Western countries have stepped up their export controls on highly toxic chemicals. The production of reagents is in a difficult situation
[0007] The application of non-toxic system to detect aflatoxin is a direction of detection technology development, and there is no report on the application of non-toxic system to detect aflatoxin by immunochromatography

Method used

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  • Test paper for testing afatoxin by nontoxic system immunochromatographic techniqul and preparing process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] A non-toxic system immunochromatographic test paper for detecting aflatoxin, comprising a bottom plate 1, a sample pad 2, a tracer particle conjugate pad 3, a nitrocellulose membrane 4, an absorbent pad 5, a detection line 6, and a quality control line 7, Among them: on the bottom plate 1, there are sample pad 2, tracer particle conjugate pad 3, nitrocellulose membrane 4, and water-absorbing pad 5 in sequence; tracer particle conjugate pad 3 has anti-aflatoxin single labeled with tracer particle For the cloned antibody, a detection line 6 and a quality control line 7 are also provided on the nitrocellulose membrane 4. The detection line 6 is composed of aflatoxin non-toxic system linear spots that can be combined with the anti-aflatoxin monoclonal antibody. The control line 7 is composed of line-like spotting of donkey anti-mouse antibody which can be combined with anti-aflatoxin monoclonal antibody.

[0103] Detection principle: Drop the sample on the sample pad 2. If ...

Embodiment 2

[0105] A method for preparing a test paper for detecting aflatoxin by immunochromatography in a non-toxic system, comprising the following steps: processing of a sample to be tested; preparation of a nitrocellulose membrane 4 detection line 6 and a quality control line 7; tracer particle conjugate pad Preparation of 3; Assembling test strips; It is characterized in that: the preparation of nitrocellulose membrane 4 detection line 6 and quality control line 7 includes the following steps: using aflatoxin non-toxic that can be combined with anti-aflatoxin monoclonal antibody The linear spotting of the system is used as the detection line 6; the linear spotting of the donkey anti-mouse antibody that can bind to the anti-aflatoxin monoclonal antibody is used as the quality control line 7. All the other are with embodiment 1.

Embodiment 3

[0107] A method for preparing a test paper for detecting aflatoxin by immunochromatography with a non-toxic system, characterized in that the non-toxic system for preparing the detection line 6 is selected in this way: 100 μl (100 μg / ml) of anti-aflatoxin monoclonal antibody is packaged at 4 ° C Block overnight with 3% skim milk at 4°C for 2h, wash 6 times with Tris-hydrochloric acid washing solution (TBST, 50mM Tris-HCl pH7.5, 150mM NaCl, 0.1% Tween20), add 100μl of phage peptide library to each well, and store at room temperature Combine for 1 hour, then wash 10 times with TBST, add 100 μl of eluent (Gly-HCL pH2.2) to each well for elution; after neutralizing the eluent, keep a part for titer determination, and add the rest to 20ml to inoculate with logarithm The E.coli ER2738 in the early growth stage was expanded and cultured in the LB culture medium. After shaking culture at 37°C for 4.5h, it was centrifuged at 10000r / min at 4°C for 10min, and 1 / 6 volume of polyethylene gl...

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Abstract

This invention relates to a method of preparing an innocuous reagent paper which is used to detect aflatoxin by immunity chromatography. It includes motherboard, sample bed, tracer grain bed, nitro cellulose membrane, water absorbing bed, detecting line, quality control line. On the motherboard, it has sample bed, tracer grain bed, nitro cellulose membrane, water absorbing bed according to priority. The tracer grain bed has anti-aflatoxin antibody marked by tracer grain. The nitro cellulose membrane has detecting line and quality control line. The detecting line is made up of aflatoxin innocuous system linear point sample, which can be combined with anti-aflatoxin antibody. The quality control line is made up of ass-anti-white rat antibody linear sample point, which can be combined with anti-aflatoxin antibody. This invention doesn't need aflatoxin in the process of produce. It avoids secondary pollution. In detect process, it doesn't need aflatoxin sample but show the result by colors. Its cost is low and it is very convenient.

Description

technical field [0001] The invention relates to a test paper for detecting aflatoxin and a preparation method thereof, in particular to a test paper for detecting aflatoxin by non-toxic system immunochromatography and a preparation method thereof. Background technique [0002] Aflatoxin is a toxin produced by Aspergillus flavus, Aspergillus parasiticus or Aspergillus model, which can cause acute poisoning, chronic poisoning, carcinogenicity, teratogenicity and mutagenicity in animals and humans. Since toxin-producing Aspergillus flavus strains widely exist in nature, it is very likely to enter the human body through the food chain, thereby posing a huge threat to human health. It is very difficult to completely avoid the pollution of aflatoxin to grain, feed, non-staple food, oil crops and fermented food. The only effective way is to strengthen the monitoring and testing of such samples, timely discover contaminated food, feed and their raw materials, and immediately remove...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558
Inventor 赖卫华熊勇华陈高明李林曾祥敏
Owner ZHONGDE DADI BIOENG JIANGXI
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