Test paper strip for rapidly detecting traces of chlorothalonil and preparation method thereof

Abstract

The invention relates to a test paper strip for rapidly detecting traces of chlorothalonil and a preparation method thereof. The test paper strip is characterized in that a supporting layer is used as the bottom layer, an adsorption layer is used as the intermediate layer, a protective layer is fixed on the adsorption layer, the adsorption layer comprises an adsorption fibrous layer, a gold-labeled antibody fibrous layer, a cellulose film and an absorbent material layer at the handle end in order from a test terminal, wherein the cellulose film is provided with detection blots printed by using a chlorothalonil coupling carrier protein solution and control blots printed by using a goat anti-mouse IgG or rabbit anti-mouse IgG (or goat anti-rabbit IgG) antibody solution; the gold-labeled antibody is a colloidal gold labeled chlorothalonil monoclonal antibody or polyclonal antibody; and the chlorothalonil coupling carrier protein is bovine serum albumin, chicken ovalbumin or haemocyanin. The test paper strip disclosed herein has the advantages of strong specificity, high sensitivity, simple, and accurate detection, low cost, wide application scope, and easiness in popularization and application.

Description

technical field

[0001] The invention relates to a device for detecting trace amounts of antibacterial drugs, in particular to a test strip for rapidly detecting trace amounts of chlorothalonil and a preparation method thereof. Background technique

[0002] Chlorothalonil (CTN), the chemical name is 2,4,5,6-tetrachloro-1,3-benzenedinitrile, the molecular formula is C 8 N 2 Cl 4 , with a molecular weight of 265.91, is a highly efficient non-systemic broad-spectrum organochlorine fungicide, which has a good preventive effect on fungal diseases of various crops, and is widely used in vegetables, fruit trees, beans, rice, wheat, forests Disease control of various crops, etc., industrially used as anti-mildew paint additives. Because chlorothalonil has good adhesion to plants, chlorothalonil and its metabolites have large residues in fruits and vegetables, soil and water, directly causing serious harm to the ecological environment and human health. At the same time, chlorothal...

Examples

preparation example Construction

[0032](1) Preparation of chlorothalonil carrier protein conjugate antigen

[0033] The carbonyl diimidazole method was used to couple chlorothalonil to carrier protein to prepare artificial conjugated antigen.

[0034] Weigh 10 mg of chlorothalonil and 3 mg of potassium fluoride and dissolve it in 400 μl of ethylene glycol solution (adding potassium fluoride can make ethylene glycol react with chlorothalonil more easily, replace the 6-position chlorine atom on the chlorothalonil molecule, and obtain half Antigen hydroxychlorothalonil (CTN-OH)), heated to 90-100°C, and magnetically stirred for 24-30 hours; after the reaction, the reaction solution was cooled to room temperature, added to 5g of ice water, and the resulting precipitate was separated and air-dried. The white solid is the chlorothalonil derivative CTN-OH (hydroxychlorothalonil); dissolve the chlorothalonil derivative with 400 μL N,N-dimethylformamide (DMF), and add 4 mg N,N-carbonyldiimidazole (CDI), magnetically ...

Embodiment 1

[0049] Example 1: For the test strips for rapid detection of trace amounts of chlorothalonil, see figure 1 , figure 2 . In the figure, the support layer 1 is made of plastic sheet strips, the absorbent fiber layer 2 is made of glass fiber cotton, the cellulose membrane layer 4 is made of nitrocellulose membrane, and the water-absorbing material layer 5 at the handle end is made of water-absorbing filter paper. 2. The gold-labeled antibody fiber layer 3, the cellulose film layer 4, and the water-absorbing material layer 5 at the handle end are pasted and fixed on the plastic sheet strip 1 in sequence from right to left, and the fibers at the junctions between the layers interpenetrate with each other. The cellulose film layer 4 is provided with a stealth detection blot 6 and a stealth control blot 7, the carrier protein coupled with chlorothalonil is bovine serum albumin (BSA), and the stealth control blot 7 is made by blotting with a rabbit anti-mouse IgG antibody. The impr...

Embodiment 2

[0055] Embodiment 2: The structure of the test strip is basically the same as that of Embodiment 1, the difference is that: the gold-labeled antibody fiber layer 3 is adsorbed with anti-chlorothalonil polyclonal antibody, the adsorption fiber layer 2 is made of nylon membrane, and the cellulose membrane Layer 4 adopts pure cellulose film, the invisible detection imprint and the invisible contrast imprint are both "ten", and 8-2 is the blue protective film on the handle end covered on the water-absorbing material layer 5.

[0056]The sample to be tested is tomato, which is cut into pieces, mashed, and then centrifuged at 4000r / min for 5 minutes, and the upper layer of juice is taken as the sample to be tested;

[0057] Result judgment: (a) Positive If there is a brown-red blot on the cellulose membrane " ten ", indicating that the test result is positive, indicating that chlorothalonil is contained in the sample to be tested; (b) negative if there are two brown-red blots on th...

technical field

[0001] The invention relates to a device for detecting trace amounts of antibacterial drugs, in particular to a test strip for rapidly detecting trace amounts of chlorothalonil and a preparation method thereof. Background technique

[0002] Chlorothalonil (CTN), the chemical name is 2,4,5,6-tetrachloro-1,3-benzenedinitrile, the molecular formula is C 8 N 2 Cl 4 , with a molecular weight of 265.91, is a highly efficient non-systemic broad-spectrum organochlorine fungicide, which has a good preventive effect on fungal diseases of various crops, and is widely used in vegetables, fruit trees, beans, rice, wheat, forests Disease control of various crops, etc., industrially used as anti-mildew paint additives. Because chlorothalonil has good adhesion to plants, chlorothalonil and its metabolites have large residues in fruits and vegetables, soil and water, directly causing serious harm to the ecological environment and human health. At the same time, chlorothal...

Examples

preparation example Construction

[0032](1) Preparation of chlorothalonil carrier protein conjugate antigen

[0033] The carbonyl diimidazole method was used to couple chlorothalonil to carrier protein to prepare artificial conjugated antigen.

[0034] Weigh 10 mg of chlorothalonil and 3 mg of potassium fluoride and dissolve it in 400 μl of ethylene glycol solution (adding potassium fluoride can make ethylene glycol react with chlorothalonil more easily, replace the 6-position chlorine atom on the chlorothalonil molecule, and obtain half Antigen hydroxychlorothalonil (CTN-OH)), heated to 90-100°C, and magnetically stirred for 24-30 hours; after the reaction, the reaction solution was cooled to room temperature, added to 5g of ice water, and the resulting precipitate was separated and air-dried. The white solid is the chlorothalonil derivative CTN-OH (hydroxychlorothalonil); dissolve the chlorothalonil derivative with 400 μL N,N-dimethylformamide (DMF), and add 4 mg N,N-carbonyldiimidazole (CDI), magnetically ...

Embodiment 1

[0049] Example 1: For the test strips for rapid detection of trace amounts of chlorothalonil, see figure 1 , figure 2 . In the figure, the support layer 1 is made of plastic sheet strips, the absorbent fiber layer 2 is made of glass fiber cotton, the cellulose membrane layer 4 is made of nitrocellulose membrane, and the water-absorbing material layer 5 at the handle end is made of water-absorbing filter paper. 2. The gold-labeled antibody fiber layer 3, the cellulose film layer 4, and the water-absorbing material layer 5 at the handle end are pasted and fixed on the plastic sheet strip 1 in sequence from right to left, and the fibers at the junctions between the layers interpenetrate with each other. The cellulose film layer 4 is provided with a stealth detection blot 6 and a stealth control blot 7, the carrier protein coupled with chlorothalonil is bovine serum albumin (BSA), and the stealth control blot 7 is made by blotting with a rabbit anti-mouse IgG antibody. The impr...

Embodiment 2

[0055] Embodiment 2: The structure of the test strip is basically the same as that of Embodiment 1, the difference is that: the gold-labeled antibody fiber layer 3 is adsorbed with anti-chlorothalonil polyclonal antibody, the adsorption fiber layer 2 is made of nylon membrane, and the cellulose membrane Layer 4 adopts pure cellulose film, the invisible detection imprint and the invisible contrast imprint are both "ten", and 8-2 is the blue protective film on the handle end covered on the water-absorbing material layer 5.

[0056]The sample to be tested is tomato, which is cut into pieces, mashed, and then centrifuged at 4000r / min for 5 minutes, and the upper layer of juice is taken as the sample to be tested;

[0057] Result judgment: (a) Positive If there is a brown-red blot on the cellulose membrane " ten ", indicating that the test result is positive, indicating that chlorothalonil is contained in the sample to be tested; (b) negative if there are two brown-red blots on th...

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