Immune detection method for residual aryl-N-methyl carbamate pesticide

A technology of methyl carbamate and esters, which is applied in the field of enzyme-linked immunoassay detection of aryl-N-methyl carbamate pesticide residues, large-scale sample detection and on-site monitoring, which can solve the complex operation and detection cost of detection methods High efficiency, low detection efficiency and other problems, to achieve the effect of good practicability, simple operation and low analysis cost

Inactive Publication Date: 2008-12-03
NANJING AGRICULTURAL UNIVERSITY
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  • Application Information

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Problems solved by technology

[0005] Technical problem The purpose of the present invention is to provide aryl-N-methyl carbamate pesticide detection method for the defects of complex operation, high detection cost and low detection efficiency of existing aryl-N-methyl carbamate pesticides. Pesticide-like multi-residue ELISA detection method, to find a fast, accurate, simple and low-cost trace detection method for aryl-N-methylcarbamate pesticide multi-residue

Method used

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  • Immune detection method for residual aryl-N-methyl carbamate pesticide
  • Immune detection method for residual aryl-N-methyl carbamate pesticide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Detection of aryl-N-methylcarbamate pesticide residues in tap water samples

[0048] Sample preparation for testing

[0049] Tap water sample: take 1L of tap water, add aryl-N-methylcarbamate pesticides A: 1 μg of standard substance of Budicarb, B: 1 μg of standard substance of Methiocarb, C: 1 μg of standard substance of Methiocarb and D: Carbaryl standard 1000μg was prepared into 1ppb, 1ppb, 1ppb and 1ppm liquid samples respectively.

[0050] The above samples were detected by ELISA detection method, and the operation was as follows:

[0051] (1) Coated

[0052] HB-OVA is used as a coating, 0.625μg / mL, add 50μL to each well of the micro-analysis plate, react in a water bath at 37°C for 2 hours, or in a refrigerator at 4°C overnight, pour it out, and wash with regular washing buffer phosphate-Tween buffer 5-7 times and inverted, pat dry on absorbent paper;

[0053] (2) closed

[0054] 1% gelatin was used as a sealer, 100 μL per well of the microanalysis ...

Embodiment 2

[0100] Example 2 Detection of Aryl-N-methylcarbamate Pesticide Residues in Small Green Vegetable Samples

[0101] Sample preparation for testing

[0102] Small green vegetables sample: Weigh 100g of dried small green vegetables, add aryl-N-methylcarbamate pesticides A: 1 μg of standard Budicarb, B: 1 μg of standard Methiocarb, C: Methiocarb Standard 1μg and D: Carbaryl standard 1000μg, prepared into 10ppb, 10ppb, 10ppb and 10ppm solid samples respectively.

[0103] The above samples were detected by the aryl-N-methylcarbamate pesticide ELISA detection method, and the operation was as follows: Steps (1) and (2) were the same as in Example 1.

[0104] (3) Add antibody and the mixture of antibody and sample to be tested

[0105] Weigh 0.2g of the sample to be tested, soak it in 0.4mL methanol for about 2 hours, take 100μL and add it to 900μL reaction matrix to prepare the test solution, monoclonal antibody (diluted by 4 times in the cell supernatant) and the test solution and b...

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Abstract

The invention relates to a method for detecting the residual immunity of an aryl-N-methylcarbamates pesticide, and belongs to the biotechnology field. The method includes the steps as follows: a carrier protein is coupled with a general hapten (HA) of the aryl-N-methylcarbamates pesticide to obtain a complete antigen; by applying the hybridoma technique, the antigen immunized Balb / C mice are screened by adopting the ELISA method and subcloned by adopting the limiting dilution method to obtain the hybridoma cell strains capable of stably secreting the aryl-N-methylcarbamates pesticide; and monoclonal antibodies are prepared. Based on the monoclonal antibodies, four ELISA detection methods for detecting the residual immunity of the aryl-N-methylcarbamates pesticide are established. The method has high specificity, sensitivity and stability, and provides a quick, sensitive and specific technology for trace-detecting the aryl-N-methylcarbamates pesticides.

Description

(1) Technical field [0001] The invention relates to an enzyme-linked immunoassay (ELISA) method for aryl-N-methylcarbamate pesticide residues, belonging to the field of biotechnology. Dedicated to the high-sensitivity and rapid detection of aryl-N-methylcarbamate pesticide residues in agricultural products and agricultural production environments, especially suitable for large-scale sample detection and on-site monitoring. (2) Background technology [0002] With the development of agricultural science and technology, pesticides have become the main means of preventing and controlling pests and diseases, and play a very important role in promoting agricultural production. Since the 1970s, due to the prohibition or restriction of organochlorine pesticides and the increasing number of insect species resistant to organophosphate insecticides, the amount of carbamate pesticides has increased year by year. At present, carbamate pesticides have become one of the most widely used p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44G01N33/577
Inventor 刘凤权孙覃胡白石张奇
Owner NANJING AGRICULTURAL UNIVERSITY
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