Colloidal gold immunochromatographic test strip for synchronously detecting diacetoxyscirpenol, deoxynivalenol and T-2 toxin

A technology of deoxynivalenol and immunochromatographic test paper, which is applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of high incidence of mycotoxins and contamination of various mycotoxins

Active Publication Date: 2020-04-14
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the scattered planting methods of small farmers in my country, the incidence of mycotoxins in agricultural products is high, and the same agricultural product is likely to be contaminated by multiple...

Method used

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  • Colloidal gold immunochromatographic test strip for synchronously detecting diacetoxyscirpenol, deoxynivalenol and T-2 toxin
  • Colloidal gold immunochromatographic test strip for synchronously detecting diacetoxyscirpenol, deoxynivalenol and T-2 toxin
  • Colloidal gold immunochromatographic test strip for synchronously detecting diacetoxyscirpenol, deoxynivalenol and T-2 toxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: Obtaining of monoclonal antibody against diacetyl sarcoenol

[0052] The anti-diacetyriferinol monoclonal antibody is produced by the secretion of the hybridoma cell line DAS5G11E7 with the preservation number CCTCC NO.C201881, and the preparation method is as follows:

[0053] The hybridoma cell line DAS5G11E7 was injected into BALB / c mice treated with incomplete Freund's adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method. Rat ascites, 4°C, centrifuge at 12000r / min for more than 15min, absorb the supernatant, mix the obtained ascites supernatant with 4 times the volume of acetate buffer, slowly add n-octanoic acid under stirring, the volume of n-octanoic acid required per milliliter of ascites 30-35μL, mix at room temperature for 30-60min, and stand at 4℃ for more than 2h. 12000r / min, centrifuge at 4°C for more than 30min, discard the precipitate, filter the obtained supernatant...

Embodiment 2

[0078] The preparation method of the immunochromatographic test strip for synchronously detecting the mixed contamination of diacetate, deoxynivalenol and T-2 toxin, the steps are as follows:

[0079] (1) Preparation of absorbent pad

[0080] Cut the absorbent paper to a length of 16mm and a width of 4mm to obtain an absorbent pad;

[0081] (2) Preparation of detection pad

[0082] The coating of the detection line:

[0083] Prepare T-2 toxin-bovine serum albumin conjugate (OTA-BSA) with coating buffer to prepare a coating solution of 0.4mg / mL, and spray it at a position 15mm away from the edge of the nitrocellulose membrane. It is coated on the nitrocellulose membrane to obtain the detection line III, and the required T-2 toxin-bovine serum albumin conjugate (the coating amount of T-2-BSA is 160ng on the detection line III per centimeter; the deoxygenated Fusarium nivalenol-bovine serum albumin conjugate (DON-BSA) was formulated with coating buffer to make a coating soluti...

Embodiment 3

[0106] The preparation method of the immunochromatographic test strip for synchronously detecting the mixed contamination of diacetate, deoxynivalenol and T-2 toxin, the steps are as follows:

[0107] (1) Preparation of absorbent pad

[0108] Cut the absorbent paper to a length of 18mm and a width of 3mm to obtain an absorbent pad;

[0109] (2) Preparation of detection pad

[0110] The coating of the detection line:

[0111] Prepare 0.5 mg / mL coating solution of diacetate sarcoenol-bovine serum albumin conjugate (DAS-BSA) with coating buffer, and place it at a position 18 mm away from the upper edge of the nitrocellulose membrane, Coat it on the nitrocellulose membrane with a dot-spraying method to obtain the detection line I, and the coating of the diacetate diacetate-bovine serum albumin conjugate (DAS-BSA) required on the detection line I per centimeter The amount is 200ng; the deoxynivalenol-bovine serum albumin conjugate (DON-BSA) is prepared into a coating solution of...

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Abstract

The invention relates to a colloidal gold immunochromatographic test strip for synchronously detecting diacetoxyscirpenol, deoxynivalenol and T-2 toxin. The test strip comprises a bottom plate. A water absorption pad, a detection pad, a gold labelled pad and a sample pad are sequentially adhered to one surface of the bottom plate from top to bottom; the adjacent pads are overlapped and connected at the joint; the detection pad takes a nitrocellulose membrane as a base pad; a quality control line and three detection lines are transversely arranged on the nitrocellulose membrane; the three detection lines are positioned under the quality control line and are distributed at intervals, the three detection lines are respectively coated with toxin protein conjugates, and the quality control lineis coated with a rabbit anti-mouse polyclonal antibody; each colloidal gold labeled antibody is transversely sprayed on the gold labeled pad; and the anti-diacetoxyscirpenol monoclonal antibody is secreted by a hybridoma cell strain DAS5G11E7 with the preservation number of CCTCC NO: C201881. The diacetoxyscirpenol, deoxynivalenol and T-2 toxin three mycotoxins can be synchronously and rapidly detected on one test strip.

Description

technical field [0001] The invention provides a colloidal gold immunochromatographic test strip and a preparation method for synchronous detection of diacetate fusarenol (serpentin), deoxynivalenol (vomitoxin), and T-2 toxin . Background technique [0002] Mycotoxins are metabolites produced by fungi growing in food or feed, and mycotoxins can be polluted during the process of planting, harvesting, storage and transportation of food and feed. Fusarenol diacetate, deoxynivalenol, and T-2 toxin are common mycotoxins in grain and feed. Fusarium acetate, also known as serpentin, belongs to the trichothecene polymycotoxins, mainly produced by Fusarium spp. and Fusarium equiseti, and deoxynivalenol and T-2 toxin are A trichothecene toxin produced by certain Fusarium fungi, three mycotoxins are found in grains including wheat, corn, barley, oats, rye, rice, millet, etc., as well as peanuts, beans, etc. Occurs widely. These three toxins are extremely harmful to humans and animal...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/531
CPCG01N33/531G01N33/558G01N33/577
Inventor 张兆威唐晓倩王督张奇李培武
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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