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Colloidal gold immunoassay test strip for synchronously detecting aspergillus flavus fungaltoxin as well as preparation and application thereof

A technology for aflatoxin and aspergillus flavus, which is applied in biological tests, analytical materials, measuring devices, etc., can solve the problem of no immunochromatographic test strips.

Active Publication Date: 2019-08-09
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no immunochromatographic test strips for various Aspergillus flavus mycotoxins such as Aspergillus versicolor and cyclopiazonic acid.

Method used

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  • Colloidal gold immunoassay test strip for synchronously detecting aspergillus flavus fungaltoxin as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Obtaining of Anti-Aflatoxin Universal Monoclonal Antibody, Anti-Varitoxin Monoclonal Antibody and Anti-Cyclopiazonic Acid Monoclonal Antibody

[0047] a. The universal anti-aflatoxin monoclonal antibody is produced by the secretion of the hybridoma cell line 1C11 with the preservation number CCTCC NO.C201013, specifically according to the method reported in the patent application number 2010102445095. The preparation method is as follows: The tumor cell line 1C11 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method. The specific operation was: filter the mouse ascites with double-layer filter paper , 4°C, centrifuge at 12000r / min for 15min, absorb the supernatant, mix the obtained ascites supernatant with 3 times the volume of acetate buffer, slowly add n-octanoic acid under stirring, the volume of n-octanoic acid require...

Embodiment 2

[0072] Colloidal gold immunoassay strips for simultaneous detection of Aspergillus flavus mycotoxins, such as figure 1 As shown, including the base plate, one side of the base plate is pasted with water-absorbing pad 1, detection pad 2, gold standard pad 3 and sample pad 4 in sequence from top to bottom. The plain film is the base pad, and the quality control line 5 and the detection line are arranged horizontally from top to bottom on the nitrocellulose membrane. The detection line is located below the quality control line, and the number is three, which are distributed at intervals. Coated with versicolor-bovine serum albumin conjugate (ST-BSA), cyclopiazonic acid-ovalbumin conjugate (CPA-OVA) and aflatoxin B1-bovine serum albumin Conjugates (AFB1-BSA), respectively Versicolor Aspergillus toxin detection line 7, cyclopiazonic acid detection line 8, aflatoxin detection line 6, the quality control line is coated with rabbit anti-mouse polyclonal antibody ; The anti-aflatoxin ...

Embodiment 3

[0101] A method for preparing a colloidal gold immunoassay strip for synchronous detection of Aspergillus flavus mycotoxins, the steps are as follows:

[0102] (1) Preparation of absorbent pad

[0103] Cut the absorbent paper into 18mm to get the absorbent pad;

[0104] (2) Preparation of detection pad

[0105] The coating of the detection line:

[0106] Prepare aflatoxin B1-bovine serum albumin conjugate with coating buffer to prepare a coating solution of 0.5 mg / mL, and spray the coating solution horizontally at a position 20 mm away from the upper edge of the nitrocellulose membrane. Coated on the nitrocellulose membrane to obtain detection line I, the coating amount of aflatoxin B1-bovine serum albumin conjugate (AFB1-BSA) required for each centimeter of detection line I was 300ng; - The bovine serum albumin conjugate was prepared into 0.5mg / mL coating solution with coating buffer, and at a position 3mm away from the detection line Ⅰ, the coating solution was horizontal...

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Abstract

The invention discloses a colloidal gold immunoassay test strip for synchronously detecting aspergillus flavus fungaltoxin as well as preparation and application thereof. The colloidal gold immunoassay test strip comprises a bottom plate, wherein an absorbent pad, a detection pad, a gold label pad and a sample pad are stuck to one side of the bottom plate from top to bottom in sequence; adjacent pads are connected at joints in an overlapping manner; the detection pad takes a nitrocellulose membrane as a base pad; a quality control line and detection lines are transversely arranged on the nitrocellulose membrane from top to bottom; three detection lines are positioned below the quality control line, and distributed at intervals; the three detection lines are coated with toxin protein conjugates respectively; the quality control line is coated with a rabbit anti-mouse polyclonal antibody; and an anti-cyclopianic acid monoclonal antibody is generated by secretion of a hybridoma cell strain YTT-2 of which the collection number is CCTCC NO.C201871. The colloidal gold immunoassay test strip can be applied to the synchronous detection of aflatoxin, sterigmatocystin and cyclopiazonic acid,is simple and rapid in operation, and has high sensitivity.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a colloidal gold immunoassay test strip for synchronous detection of aspergillus flavus mycotoxins, its preparation and its application. Background technique [0002] Aspergillus flavus is a common saprophytic aerobic fungus, which exists in the form of conidia or sclerotia in the soil, and in the form of mycelium in plant tissues. The sclerotia can survive under extreme environmental conditions (high temperature, drought, etc.) to survive. Aspergillus flavus has multiple secondary metabolic gene clusters, which can produce toxic secondary metabolites such as aflatoxin, aspergillus versicolor and cyclopiazonic acid, which can induce a variety of important agricultural products before and after harvesting, such as The occurrence of diseases such as rice, corn, peanuts, and feed causes the contamination of Aspergillus flavus mycotoxins. Aflatoxin is a class I carci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577G01N33/58G01N33/532
CPCG01N33/558G01N33/577G01N33/58G01N33/532
Inventor 张奇李培武王督张兆威张文
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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