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Colloidal gold immune chromatography test paper for detecting biotoxin and detecting method thereof

A technology for immunochromatographic test strips and biological toxins, which is applied in measurement devices, analytical materials, instruments, etc., can solve the problems of expensive instruments, long time required, poor specificity, etc., and achieves simple and convenient operation, wide application area and specificity. strong effect

Active Publication Date: 2008-10-08
CHINA JILIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have certain limitations: the biological detection method is relatively simple and low in cost, but it takes a long time and has poor specificity; enzyme-linked immunoassay (ELISA) is currently recognized as a method for the determination of mycotoxin One of the best methods (publication numbers are: CN1963506A, CN1877332A Chinese invention patent applications), high sensitivity, strong specificity, no need for expensive instruments and equipment, low requirements for sample purity, suitable for the detection of large batches of samples, but this method It still needs to be completed in the laboratory, which takes 1-2 hours, and cannot meet the needs of on-site rapid detection; physical and chemical detection methods mainly include thin-layer chromatography, high-performance liquid chromatography, gas chromatography, and chromatography-mass spectrometry, etc. However, thin-layer chromatography has low detection sensitivity and requires a large amount of contact with standard substances, which is not conducive to protecting the health of operators. High-performance liquid chromatography, gas chromatography, and chromatography-mass spectrometry are sensitive and accurate, but expensive instruments and samples The processing is cumbersome, time-consuming, costly, and requires professional operation

Method used

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  • Colloidal gold immune chromatography test paper for detecting biotoxin and detecting method thereof
  • Colloidal gold immune chromatography test paper for detecting biotoxin and detecting method thereof
  • Colloidal gold immune chromatography test paper for detecting biotoxin and detecting method thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0048] (2) Preparation of spleen cell suspension:

[0049] Mice with high titers were killed by pulling their necks, soaked in 70% alcohol for 10 minutes, and then cut a small opening in the abdomen of the mice with surgical scissors, peeled off the skin, exposed the abdominal cavity, and took out the spleen under sterile conditions. The solution was washed once, placed on a stainless steel mesh in a dish, ground into a cell suspension with a syringe needle and counted.

[0050] (3) Cell fusion and cloning:

[0051] Take the logarithmically growing myeloma cells, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, suspend the cells with the incomplete medium and count them, take the required number of cells, and wash twice with the incomplete medium. At the same time, a suspension of immune spleen cells was prepared and washed twice with incomplete culture medium. Myeloma cells and splenocytes were mixed together at a ratio of 1:10 or 1:5, washed once with incompl...

Embodiment 1

[0076] Example 1 Quadruple detection test paper for DON, zearalenone, ochratoxin A and fumonisin B1:

[0077] The test strip detection line is coated with an antigen for biotoxin detection at a concentration of 0.5-2.0 mg / mL, such as figure 2 As shown, T4 is 4-6mm from the bottom of the detection area; the quality control line is coated with goat anti-mouse IgG with a concentration of 0.5-1.5mg / mL, and it is 4-6mm from the top of the detection area; C line and T1, T1 and T2, T2 With T3, the interval between T3 and T4 is 2-4mm.

[0078] Test result judgment:

[0079] 1. If figure 2 The test lines T1, T2, T3, T4 and quality control line C shown in the middle A are all red, indicating that the test result is negative, indicating that DON, zearalenone, ochratoxin A and fumonis are in the tested sample. The contents of the three biotoxins of B1 were below the detection limit.

[0080] 2. If figure 2 The test lines T1, T2, T3 and quality control line C shown in middle B are ...

Embodiment 2

[0118] Example 2 Test paper for triple detection of DON, zearalenone and fumonisin B1:

[0119] The test strip detection line is coated with an antigen for biotoxin detection at a concentration of 0.5-2.0 mg / mL, such as Figure 4 As shown, T3 is 6-8mm from the bottom of the detection area; the quality control line is coated with goat anti-mouse IgG with a concentration of 0.5-1.5mg / mL, and it is 6-8mm from the top of the detection area; C line and T1, T1 and T2, T2 The interval between the two lines of T3 is 2-4mm.

[0120] Test result judgment:

[0121] 1. If Figure 4 The test lines T1, T2, T3 and quality control line C shown in middle A are all red, indicating that the test result is negative, indicating the content of three biological toxins, DON, zearalenone and fumonisin B1 in the tested sample below the detection limit.

[0122] 2. If Figure 4 The test lines T1, T2 and quality control line C shown in middle B are red, and the test line T3 is not colored. The test ...

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Abstract

The invention pertains to the biotoxin testing technique field, especially to a colloidal gold immunity chromatography reagent paper for testing biotoxin and testing method thereof. The colloidal gold immunity chromatography reagent paper for testing biotoxin includes a colloidal gold marking mat and testing wires, wherein, the substance covered by the colloidal gold marking mat may be a mixer ofcolloidal gold marking articles of a biotoxin antibody or a plurality of biotoxin antibodies, the number of the testing wires is corresponding to the number of the biotoxin antibodies covered by the colloidal gold marking articles, and each testing wire is cover with a single biotoxin testing antigen corresponding to the colloidal gold marking antibody. The inventive colloidal gold immunity chromatography reagent paper has strong specificity; has convenient and simple operation, which can direct detect milk and animal urine, can test samples such as feed, wheat, corn, barley and the like after simple processing, and can observe result 3-5min later. The inventive method has wide application, can satisfy requirements of food product safety, feed safety as well as fast detecting of biotoxin residual by governmental detection mechanism.

Description

technical field [0001] The invention belongs to the technical field of biological toxin detection, in particular to a colloidal gold immunochromatographic test paper for detecting biological toxins and a detection method thereof. Background technique [0002] Biotoxins are anti-nutritional factors present in feedstuffs and feedstuffs, and are secondary toxic metabolites produced by different species of fungi. Contamination of biotoxins in animal feed is a global problem. Biotoxins can be produced during crop field growth, during storage, or both. Feeding animals with biotoxin-contaminated feeds can cause acute and chronic The symptoms of poisoning can even lead to death, and it will remain in the meat, eggs and milk of animals, threatening the health of human beings. [0003] The main biotoxins in animal feed are mycotoxins, which are produced by mesophilic molds. The common toxin-producing molds in grains and feeds are: Aspergillus, Penicillium, Fusarium ( fusarium) and e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577G01N33/531
Inventor 张明洲陈宗伦刘军郭瑞忠施明华胡华军储国华
Owner CHINA JILIANG UNIV
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