Quantitative rapid deoxynivalenol detection card and detection method thereof

A technology of deoxynivalenol and detection card, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problem of affecting the accuracy and stability of test results, difficult to distinguish negative and positive test results, and difficult to be accurate in particle size deviation. Control and other issues, to achieve the effect of saving manpower and material resources, easy to distinguish, and small differences

Inactive Publication Date: 2018-11-06
HEBEI ELISHA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] (1) Colloidal gold itself is dark purple red, and the color is relatively light, so it is difficult to distinguish the positive and negative of the test results;
[0006] (2) The amount of gold produced in one batch of colloidal gold is at most about 10L, and the particle size deviation is difficult to accurately control. Quantitative products are mass-produced, and different batches are likely to cause large batch-to-batch differences, which will affect the accuracy and stability of the test results;
[0007] (3) Colloidal gold labeling techn

Method used

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  • Quantitative rapid deoxynivalenol detection card and detection method thereof
  • Quantitative rapid deoxynivalenol detection card and detection method thereof
  • Quantitative rapid deoxynivalenol detection card and detection method thereof

Examples

Experimental program
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Embodiment 1

[0060] to combine figure 1 As shown, a deoxynivalenol quantitative rapid detection card provided in this embodiment includes a sample pad 2, a nitrocellulose membrane 3 and a water-absorbing pad 4 arranged on the base plate 1 in sequence, and does not include colloidal gold Release pad of labeled antibody or monoclonal antibody with detection line (T line) coated with deoxynivalenol hapten-carrier protein coupling agent (DON-BSA) on the nitrocellulose membrane 3 5 and the quality control line (C line) 6 coated with goat anti-mouse antibody, the sample solution to be tested is mixed and incubated with latex microspheres coupled with DON antibody before being dropped onto the sample pad.

[0061] In this embodiment, the coating amounts of the detection line 5 and the quality control line 6 are respectively 0.1 mg / mL and 0.4 mg / mL, and the nitrocellulose membrane 3 is After the coating of the quality control line 6, it was dried overnight at 37°C.

[0062] In this embodiment, ...

Embodiment 2

[0074] The deoxynivalenol quantitative rapid detection card described in this example is the same as in Example 1, except that the DON antibody is a monoclonal antibody, and the monoclonal antibody is prepared by the following method:

[0075] (1) Preparation of immunogen

[0076] Dissolve 8 mg of deoxynivalenol hapten in 0.8 mL of DMF, then add 20 mg of EDC fully dissolved in 0.2 mL of water, and stir at room temperature for 24 hours to obtain reaction solution A; weigh 36 mg of OVA was fully dissolved in 3 mL of CB with a concentration of 0.1 mol / L and a pH of 9.6 to obtain a protein solution A; the reaction solution A was slowly added dropwise to the protein solution A, and stirred at room temperature for 24 Hours, dialyze with PBS with a concentration of 0.01mol / L at 4°C for 3 days and change the dialysate 3 times a day to obtain the immunogen;

[0077] (2) immunity

[0078] Immunizing Balb / c mice with the immunogen obtained in step (1), the immunological dose is 100 μg / ...

Embodiment 3

[0086] to combine figure 2 As shown, the present embodiment provides a kind of detection method of deoxynivalenol quantitative rapid detection card described in embodiment 1, comprises the steps:

[0087] (1) Preheating: turn on the incubator switch and raise the temperature to 45±1°C before testing;

[0088] (2) Labeled samples: According to the number of samples to be tested, take out the corresponding number of latex microspheres coupled with DON antibody and the test card that have been lyophilized and coated, and mark on the test card to distinguish the sample solution to be tested

[0089] (3) Liquid addition: use a pipette to draw 120ul of the sample solution to be tested, drop it vertically into the latex microspheres coupled with DON antibody that have been lyophilized and coated, and repeatedly pipette for more than 10 times until the mixture is uniform;

[0090] (4) Incubation: place the detection card and the solution obtained in step (2) on the incubator, and inc...

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Abstract

The invention discloses a quantitative rapid deoxynivalenol detection card which comprises a sample pad, a nitrocellulose membrane and an absorbent pad arranged on a bottom plate in sequence, whereina detection line coated with a DON hapten-carrier protein coupling agent and a quality control line coated with a goat anti-rat antibody are arranged on the nitrocellulose membrane; a to-be-detected sample solution is mixed and incubated with latex microspheres coupled with a DON antibody before dropping to the sample pad. The detection method comprises the steps: preheating, adding liquid, incubating, reacting and reading the result. The detection card disclosed by the invention does not need a release pad containing a gold-labelled antibody and is used for detecting the latex microspheres coupled with the DON antibody mixed with the sample solution, the color is bright and easy to determine, and the coupling antibody is in a chemical bond coupling state, has excellent physical adsorptionstability with colloidal gold and is high in detection accuracy and precision degree; the batch-to-batch difference is small, and the cost is low; in addition, the detection method of the detection card disclosed by the invention is simple in operation and suitable for batch production and application.

Description

technical field [0001] The present invention relates to a quantitative rapid detection card and its detection method, in particular to a deoxynivalenol quantitative rapid detection card and its detection method, belonging to the technical field of biological detection, especially suitable for grain, oil and grain Quantitative and rapid detection of a class of deoxynivalenol. Background technique [0002] Vomitoxin (vomitoxin), also known as deoxynivalenol (deoxynivalenol, DON), chemical name 3α, 7α, 15-trihydroxy Fusarium-9-en-8-one, belongs to Trichothecenes mycophenolic compounds. It gets its name because it can cause pigs to vomit, and it also has certain harmful effects on the human body. The EU classification standard is a third-class carcinogen. Vomitoxin is highly toxic to both humans and animals, and can cause vomiting, diarrhea, skin irritation, food refusal, nervous disorders, miscarriage, stillbirth, etc. Pigs are the most sensitive animals to vomitoxin, followe...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543
CPCG01N33/54306G01N33/54313G01N33/56911
Inventor 刘建龙吴文娟王俞丹
Owner HEBEI ELISHA BIOTECH CO LTD
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