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Enzyme-linked immunosorbentassay reagent kit for analyzing snout-killing sulfur-phosphous residual

An enzyme-linked immunosorbent adsorption and fenitrothion technology, applied in the direction of analysis materials, measuring devices, instruments, etc., can solve the problems of long time-consuming, complicated and expensive operation process, and achieve less time-consuming, simple pre-treatment process, and improved The effect of sensitivity

Inactive Publication Date: 2005-10-05
韩丽君 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the analysis of pesticide residues, the conventional detection method for fenitrothion is gas chromatography, but this method requires cumbersome pretreatment steps such as extraction, purification, and concentration of the sample, and this method requires expensive chromatographic instruments, and the operation process is complicated And it takes a long time, it is not suitable for the detection and analysis of large batches of samples

Method used

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  • Enzyme-linked immunosorbentassay reagent kit for analyzing snout-killing sulfur-phosphous residual
  • Enzyme-linked immunosorbentassay reagent kit for analyzing snout-killing sulfur-phosphous residual
  • Enzyme-linked immunosorbentassay reagent kit for analyzing snout-killing sulfur-phosphous residual

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Effect test

Embodiment 1

[0026] The product of the present invention includes a box body, a 96-well / 40-well microplate plate in the box body, and detection reagents. In each well of the microplate plate, the coating solution that can specifically bind to the anti-fenitrothion antibody is coated Antigen, and 2% to 5% gelatin for blocking.

[0027] Reagents in the box include: washing solution (diluent), substrate diluent, anti-fenitrothion antibody (suitable for indirect ELISA), fenitrothion standard solution, horseradish peroxidase-labeled goat anti-rabbit antibody ( Suitable for indirect competition ELISA) or horseradish peroxidase-labeled anti-fenitrothion rabbit antibody (for direct competition ELISA), substrate, chromogenic substance and reaction termination solution, the preparation method is as follows:

[0028] Washing solution (reaction diluent) 40mL, the composition ratio is 0.1g of potassium dihydrogen phosphate, 4g of disodium hydrogen phosphate, 0.1g of potassium chloride, 3mL of Tween-20,...

Embodiment 2

[0036] Preparation of enzyme-labeled antibody (improved sodium periodate method), the specific operation is as follows:

[0037] Weigh 5-10mg of horseradish peroxidase HRp and dissolve it in 1L of distilled water, add 0.2-04mL of freshly prepared 0.1mol / L NalO4 solution, and stir for 15-30min at room temperature in the dark. Put the above solution into a dialysis bag, dialyze with 1 mmol / L acetate buffer solution of pH 4.4, overnight at 4°C. Use a carbonate buffer solution of pH 9.5 to raise the pH of the above hydroformylated HRP solution to 9.0-9.5, then immediately add 1-2 mL of a 0.01 mol / L carbonate buffer solution containing 10-20 mg of purified antibody, and store at room temperature Stir in the dark for 2-3 hours. Add 0.1-0.2mL of freshly prepared 4mg / mL NaBH4 solution, mix well, and place at 4°C for 2-3 hours. The reaction solution was put into a dialysis bag, dialyzed with 0.15 mol / L PBS, pH7.4, overnight at 4°C. Add an equal volume of saturated ammonium sulfate s...

Embodiment 3

[0039] Preparation of Coated ELISA Plates

[0040] Coating antigen (FEN-OVA) was diluted to 0.5-4 μg / mL with pH 9.6 0.05 mol / L carbonate buffer solution (containing 1-2 g sodium carbonate and 2-4 g sodium bicarbonate, 1 L double-distilled water), Add 100 μL to each well of the ELISA plate, coat overnight at 4°C or 2 hours at 37°C, pour off the coating solution, wash 3 times with PBST, pat dry, and then add 150 μL of 2%-5 % gelatin, placed in a 37°C incubator to seal for 0.4 to 1 hour, washed 3 times with PBST, patted dry and stored in a dry place.

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Abstract

The invention discloses a enzymoimmunoassy absorption test agent box adapted on fenitrothion remain analysis which includes a box, 96 hole or 40 hole enzyme labeling board or test tube set in the box and the antigen in the box. In each hole on the enzyme labeling board, envelope antigen which is able to unite with fenitrothion antigen, is enveloped by parisarc liquor, and then sealed with celluloid of 2 percents to 5 percents. Antigens in the box consists of washing solution, basal material dilution, fenitrothion standard solution, fenitrothion antigen (indirect ELISA), sheep rabbit antigen labeled with horse radish peroxidase (indirect ELISA), or fenitrothion rabbit antigen labeled with horse radish peroxidase (direct ELISA), basal material, coloration matter and reaction termination liquor. The invention can be used on fast detecting of fenitrothion remain in water, soil, vegetable, fruits and poisoned sample. The pre-process of the sample is simple and can detecting groups of samples at the same time. The sample detecting cost is lower than traditional detecting method.

Description

(1) Technical field [0001] The invention belongs to an enzyme-linked immunosorbent assay kit suitable for analyzing residual fenitrothion. The kit is mainly suitable for rapid determination of batches of environmental samples such as water samples, soil, poisoned samples, and food samples such as vegetables. Fenitrothion residues belong to the field of pesticide residue detection. (2) Background technology [0002] Due to the continuous development of pesticide use technology and the continuous expansion of the scale of use, the environmental impact caused by pesticide residues and the chronic and long-term effects on human health are increasingly concerned and worried, and the restrictions on pesticide residues in daily consumption and foreign trade are also increasing. It is becoming more and more strict, and puts forward new requirements and higher standards for the analysis and detection of objects, types, quantities, scope, indicators and many other aspects. In terms o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/547
Inventor 韩丽君钱传范
Owner 韩丽君
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