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Enzyme-linked immune analytic method for detecting carbofuran

An enzyme-linked immunoassay and carbovir technology, applied in the field of enzyme-linked immunoassay, can solve the problems of expensive professional instruments, time-consuming analysis, and high cost, and achieve the effects of high affinity, low cost, and reduced inhibition rate.

Inactive Publication Date: 2007-06-27
SOUTH CHINA AGRI UNIV
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AI Technical Summary

Problems solved by technology

[0003] At present, the conventional methods for detecting carbofuran residues include gas chromatography (GC) and high performance liquid chromatography (HPLC). Although these methods are sensitive and accurate, they require expensive professional instruments, and the analysis is time-consuming and costly.

Method used

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  • Enzyme-linked immune analytic method for detecting carbofuran

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Embodiment 1

[0021] 1. Preparation of buffer buffer (pH7.4PBST) KH 2 PO 4 0.4g, Na 2 HPO 4 12H 2 O5.8g, NaCl 16g, KCl 0.4g, Tween-200.05% 1mL, add distilled water to 2000mL.

[0022] 2. Preparation of blocking solution Dissolve 3.0 g of skimmed milk powder in 100 mL of distilled water.

[0023] 3. Preparation of substrate solution Dissolve 30 μL of 30% hydrogen peroxide in 19 mL of chromogenic solution (pH 5.0 phosphoric acid-citric acid buffer 0.2 M Na 2 HPO 4 25.7mL, 0.1M citric acid 24.3mL, add distilled water 50mL), store at 4°C.

[0024] 4. Preparation of substrate buffer solution Dissolve 80 mg o-phenylenediamine OPD in 10 ml substrate solution and store at 4°C.

[0025] 5. Dilute the coated secondary antibody of the microplate to 4ug / ml with pH9.6, 0.05mol / L carbonate buffer solution (containing 2g sodium carbonate and 4g sodium bicarbonate, 1L of double distilled water), and in the enzyme Add 100ul to each well of the target plate, coat overnight at 4°C, pour off the coat...

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Abstract

Enzyme immunoassay method for detecting Kebaiwei includes (1) synthesizing half antigen and artificial antigen; (2) preparing polyclonal antibody, monoclonal antibody, or gene engineering antibody of possessing specificity to Kebaiwei; (3) preparing enzyme labeled antigen; (4) using second antibody to coat enzyme labeled plate and close the plate to save Kebaiwei; reaction is carried out between anti Kebaiwei antibody and second antibody and fixed on the enzyme labeled plate; (5) washing out free object, adding sample to be tested, and enzyme labeled antigen or Kebaiwei labeled sample, and enzyme labeled antigen; (6) washing out free object, adding substrate of enzyme and developer. Intensity of enzyme promoting color reaction is proportional to quantity of enzyme labeled antigen bound on antibody, and is inversely proportional to content of Kebaiwei in sample. Features are: long term storage, increasing sensitivity, accuracy, simple and quick test operation.

Description

technical field [0001] The invention relates to an enzyme-linked immunoassay method, in particular to an enzyme-linked immunoassay method for detecting carbofuran. Background technique [0002] Carbofuran (carbofuran, chemical name 2,3 dihydro-2,2 dimethyl-7-benzofuryl-N-methylcarbamate), trade name carbofuran, has been produced by FMC and After being developed and produced by Mobay Chemical Company, it is used as a high-efficiency and broad-spectrum insecticide, which is widely used in the control of pests in grains, vegetables, fruits and economic crops. Carbofuran is a cholinesterase inhibitor that is highly toxic to humans and wild animals (oral LD ​​in mice 50 8mg / kg), it is a kind that cannot be detected in the regulations on pesticide residues in vegetables, but due to its good insecticidal effect, the phenomenon of unreasonable use in food and vegetables is relatively serious, and it has great harm to public health . In addition, carbofuran is not easy to degrade ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/547G01N21/78G01N33/53
Inventor 孙远明杨金易潘科王弘吴青雷红涛肖治理谌国莲沈玉栋
Owner SOUTH CHINA AGRI UNIV
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