Enzyme-linked immunosorbant assay kit applicable for carbofuran residue analysis

An enzyme-linked immunosorbent adsorption and residue analysis technology, applied in the direction of analytical materials, biological testing, measuring devices, etc., can solve the problems of cumbersome process, great influence on sensitivity, unsuitable for large-scale sample detection and analysis, etc., to improve sensitivity, The effect of simple pre-processing and less time-consuming

Inactive Publication Date: 2002-12-18
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the sensitivity of this method is greatly affected by the steps of sample purification, concentration, derivatization, etc.
Furthermore, this method requires complex instrumen

Method used

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  • Enzyme-linked immunosorbant assay kit applicable for carbofuran residue analysis
  • Enzyme-linked immunosorbant assay kit applicable for carbofuran residue analysis

Examples

Experimental program
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Effect test

Embodiment 1

[0011] The principle of determination is that first, the compound obtained 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 corresponding antibody are added, and the pesticide on the solid-phase antigen , the pesticide to be tested competes with the antibody. If the pesticide to be tested contains more, the antibody bound to the solid-phase antigen will be less. On the contrary, the antibody bound to the solid-phase antigen will be more. After the reaction, add an enzyme-labeled secondary antibody (which can only be combined with Combined with the antibody on the solid-phase antigen), and finally use the substrate for color development to determine. When the amount of antibody is constant, the more the amount of pesticide to be tested is added, the less the antibody combined with the solid-phase antigen, and the color reaction If it is we...

Embodiment 2

[0013] For the preparation of coated microtiter plates, the coated antigen (BFNB-OVA or BFNH-OVA) was coated with pH9.6, 0.05mol / L carbonate buffer solution (containing 1-2g sodium carbonate and 2-4g sodium bicarbonate, Double-distilled water (1 L) diluted to 0.1-1 μg / mL, add 100 μL to each well of the microtiter plate, coat overnight at 4°C or 2h at 37°C, pour off the coating solution, wash 3 times with PBST, and pat dry , and then add 150 μL of 1.0-3.0% skimmed milk powder to each well, put it in a 37°C incubator for 0.4-1 hour, wash with PBST three times, pat dry and store in a dry place. Example 3

Embodiment 3

[0014] Pretreatment of test samples:

[0015] Water sample: After filtration, samples can be taken for ELISA analysis.

[0016] Soil sample: Take 10g of soil and extract it three times with 20-40mL of methanol, combine the extracts, concentrate, then dilute with PBST to 10mL for ELISA analysis.

[0017] Vegetable samples: Take vegetable samples and crush them with a pulverizer, weigh 10g, extract three times with 20-40mL of methanol, combine the extracts, concentrate, dilute to 10mL with PBST, and take samples for ELISA analysis.

[0018] Blood: take human blood, add anticoagulant and directly analyze it by ELISA method.

[0019] Gastric lavage solution (2% sodium bicarbonate solution): take 10 mL of gastric lavage solution, adjust the pH value to neutral with dilute HCl, and then analyze it by ELISA.

[0020] Vomitus: Take the sample and grind it, centrifuge to get the supernatant and analyze it by ELISA method.

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Abstract

The invention discloses a kind of enzyme linked immuno-sorbent assay reagent box applying to the analysis of the Carboruran residue, and it includes the box body, the 96-hole/40-hole enzyme marker board placed in the box body and the reagent placed in the box body, and its characteristic lies in that in each hole of the enzyme marker board is the envelope antigen enveloped by the envelope liquid and able to combined and react with the anti-Carboruran antibody specioficity and sealed with 1.0-3.0% nonfat dried milk, and that the reagent in the box includes cleaning mixture, substrate-matter diluents, Carboruran standard solution, anti-CArboruran antibody, horseradish peroxide enzyme marker sheep anti-rabbit antibody, substrate matter, color-displaying matter and reaction-termiinated liquor.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay kit suitable for the analysis of carbofuran residues, which is mainly suitable for the rapid determination of carbofuran in large quantities of vegetables, water, soil, poisoning and other environments, food and other samples by using the kit residual. 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 traditional...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/535G01N33/543
Inventor 朱国念吴慧明杨挺程敬丽
Owner ZHEJIANG UNIV
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