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Test kit of enzyme linked immuno sorbent assay applicable to methylparathion retention analysis

An enzyme-linked immunosorbent assay, methyl parathion technology, applied in analytical materials, biological tests, measurement devices, etc., to achieve the effect of less time-consuming, simple pretreatment process, and improved sensitivity

Inactive Publication Date: 2003-07-23
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Test kit of enzyme linked immuno sorbent assay applicable to methylparathion retention analysis
  • Test kit of enzyme linked immuno sorbent assay applicable to methylparathion retention analysis
  • Test kit of enzyme linked immuno sorbent assay applicable to methylparathion retention analysis

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Experimental program
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Effect test

Embodiment 1

[0012] The principle of its determination is that firstly, the compound prepared by coupling the pesticide molecule with the macromolecular carrier (such as protein) is adsorbed on the solid phase carrier as the coated antigen, and then the pesticide to be tested and the enzyme-labeled antibody are added, and the concentration on the solid phase antigen Pesticide, the pesticide to be tested competes with the enzyme-labeled antibody. If the content of the pesticide to be tested is large, the enzyme-labeled antibody bound to the solid-phase antigen is less. On the contrary, the enzyme-labeled antibody bound to the solid-phase antigen is more, and the substrate is added after the reaction Carry out color development to determine, when the amount of enzyme-labeled antibody is constant, the more the amount of pesticide to be tested is added, the less the enzyme-labeled antibody combined with the solid-phase antigen, the color reaction is weakened, and the inhibition rate is increased...

Embodiment 2

[0014] Preparation of enzyme-labeled antibody (using the modified sodium periodate method)

[0015] The specific operation is as follows: Dissolve 5-10 mg of HRP in 1 mL of distilled water, and add 0.2-0.4 mL of newly prepared 0.1 mol / L NaIO to the supernatant 4 The solution was stirred at room temperature in the dark for 15-30 minutes. Put the above solution into a dialysis bag, dialyze with 1 mmol / L acetate buffer solution of pH 4.4, overnight at 4°C. Add 20-40 μl of 0.2mol / L pH9.5 carbonate buffer to raise the pH of the above hydroformylated HRP to 9.0-9.5, then immediately add 1-2ml of 0.01mol / L carbonate buffer containing 10-20mg of purified antibody Salt buffer solution, stirred gently at room temperature for 2 to 3 hours in the dark. Add 0.1~0.2mL of newly prepared 4mg / mL NaBH 4 solution, mix well, and place at 4°C for 2 to 3 hours. The reaction solution was put into a dialysis bag, and dialyzed with 0.15mol / L pH7.4 PBS at 4°C overnight. Add an equal volume of satu...

Embodiment 3

[0017] Preparation of Coated ELISA Plates

[0018] Coating antigen (M1605-OVA) was diluted with pH9.6, 0.05mol / L carbonate buffer solution (containing 1-2g sodium carbonate and 2-4g sodium bicarbonate, 1L of double distilled water) to 0.5-4μg / mL , add 100 μL to each well of the ELISA plate, coat overnight at 4°C or 2h at 37°C, pour off the coating solution, wash 3 times with PBST, pat dry, and then add 150 μL of 1.0-3.0 % skimmed milk powder, placed in a 37°C incubator for 0.4 to 1 hour, washed with PBST three times, patted dry and stored in a dry place.

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Abstract

The test kit includes the box body, the 96-eyelets enzyme target / test tube in the box, and the reagent in the box. In each eyelet of the enzyme target, the peridium fluid coats the peridium antigen, which generates the specificity fixatino reaction with anti metacide antibody and the 1.0-3.0% defatted milk powder used to carry out the closing. The reagent includes the cleaning solution, the diluentt, and standard metacide fluid, the anti metacide antibody, the horseradish peroxidase marked rabbit antibody of anti metacide, the substrate, the coloration matter and the reaction termination fluid. The invention provides the fast testing, the simple procedure for preprocessing the sample, not time-consuming, and capable of testing in batch mode.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay kit suitable for the analysis of methyl parathion residues, which is mainly suitable for rapid determination of batches of water samples, soil and other environmental samples, poisoned samples, and food samples such as vegetables. of methyl parathion residues. Background technique [0002] The traditional residue analysis methods of pesticides and their metabolites mainly rely on physical and chemical analysis methods such as gas chromatography (GC), high performance liquid chromatography (HPLC), and mass spectrometry (MS). The chronic and long-term effects of human health and human health are increasingly concerned and worried, and the restrictions on pesticide residues are becoming more and more stringent. New requirements and higher requirements are put forward for the analysis and measurement objects, types, quantities, ranges, indicators, etc. However, the tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/535G01N33/558G01N33/573
Inventor 吴慧明程敬丽魏方林杨挺吴刚朱国念
Owner ZHEJIANG UNIV
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