Reagent kit and enzyme-linked immunochromatography for detecting various organophosphorus pesticide residues

An organophosphorus pesticide and enzyme-linked immunosorbent technology, applied in measuring devices, analytical materials, instruments, etc., can solve the problem of low sensitivity and achieve high sensitivity, good resolution, and intuitive results

Inactive Publication Date: 2012-07-04
SHENZHEN KANGMEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The technical problem to be solved in the present invention is to provide an ELISA with high sensitivity, strong specificity, convenient and quick detection of

Method used

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  • Reagent kit and enzyme-linked immunochromatography for detecting various organophosphorus pesticide residues
  • Reagent kit and enzyme-linked immunochromatography for detecting various organophosphorus pesticide residues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Quantification of organophosphorus pesticides after preparation of enzyme-labeled antibody by improved sodium periodate method

[0042] (1) Preparation of enzyme-labeled antibody by improved sodium periodate method

[0043] Dissolve 5mg of HRP in 0.2mol / L, pH5.6 acetate buffer 1ml, add 0.1ml of 1% DNFB absolute ethanol solution, stir gently at room temperature for 1h; add freshly prepared 0.1mol / L NaIO 4 0.5ml, place at 4°C for 30min, add 1ml of 2.5% ethylene glycol, stir gently at room temperature for 1h, and terminate the reaction; add 5-10mg of the antibody to be labeled, and adjust the pH value to 9.0; mix well, put at 4°C overnight; add 0.1ml of sodium borohydride solution, mix evenly, place at 4°C for 3 hours; dialyze against 0.01mol / L, pH7.4 PBS overnight at 4°C, change the solution 3 times; 3000r / min Centrifuge for 30 minutes to remove the precipitate; collect the supernatant to be the enzyme-labeled antibody.

[0044] (2) Test strip assembly

[0...

Embodiment 2

[0055] Example 2: Quantitative detection of organophosphorus pesticides after labeling antibodies with biotin-avidin combined with ELISA

[0056] (1) Preparation of biotin-labeled antibody.

[0057] The antibody was dialyzed against 50 mM carbonate buffer (pH 9.6) at 4°C to remove sodium azide (NaN3), and the anti-organophosphorus monoclonal antibody to be biotinylated was diluted to 1 mg / ml with the carbonate buffer. Biotin ester (BNHS) was dissolved in dimethylformamide (DMF) at a concentration of 20 mg / mL. Add BNHS at the ratio of 100 μg BNHS per 1 mg antibody, mix well, and react at room temperature for 4 hours. Dialyze with phosphate buffered saline for three days at 4°C to remove free matter, and obtain biotinylated monoclonal antibody, which is stored at 4°C for later use, and stored at -20°C for long-term storage.

[0058] (2) Assembly of test strips

[0059] The test strips were assembled according to the method of Example 1.

[0060] (3) Principle of detection pr...

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Abstract

The invention discloses a reagent kit and an enzyme-linked immunochromatography for detecting various organophosphorus pesticide residues and particularly relates to the reagent kit and the enzyme-linked immunochromatography for detecting organophosphorus pesticide residues in the production of vegetative agricultural products. The enzyme-linked immunochromatography for detecting the organophosphorus pesticide residues uses the high-sensitivity characteristic of enzyme to carry out detection by adopting the immunochromatography; compared with a traditional enzyme-linked immunoserbent assay (ELISA) for detecting organophosphorus pesticides, the enzyme-linked immunochromatography has the advantages of simplicity and convenience in operation, and visual result, and no special instrument equipment is needed; and compared with a colloidal gold immunochromatography, the enzyme-linked immunochromatography has the advantages of high sensitivity, strong specificity, accuracy in quantification by combining with a readout instrument and the like.

Description

technical field [0001] The invention relates to the technical field of pesticide detection, in particular to an enzyme-linked immunochromatographic method and a kit for detecting residues of various organophosphorus pesticides. Background technique [0002] Pesticide residues refer to the general term for the trace pesticide precursors, toxic metabolites, degradation products and impurities remaining in organisms, food (agricultural by-products) and the environment after pesticide use. Pesticides pollute food through direct or indirect routes (atmosphere, water, soil). Human beings are at the top of the food chain, and suffer the most serious harm from bioaccumulation of pesticide residues. Today, pesticide residues have become a major food safety issue in Chinese diets. Pesticides that seriously pollute food are mainly organophosphorus, organochlorine, carbamate and pyrethroid insecticides, fungicides and herbicides. There are about 500 kinds of pesticides commonly used ...

Claims

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Application Information

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IPC IPC(8): G01N33/558
Inventor 王金爱
Owner SHENZHEN KANGMEI BIOTECH
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