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Pesticide ethofenprox fluorescence quantitative immunochromatography test paper and application thereof

A technology for immunochromatographic detection and fluorescence quantification, which can be used in the biological field to solve problems such as sensitivity gaps

Active Publication Date: 2020-10-16
EXPERIMENTAL RES CENT CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, colloidal gold immunochromatography (gold standard test paper), as a representative detection method, has the advantages of on-site, fast, no need for equipment, and can directly judge the results. It is a quantitative or semi-quantitative method, and its sensitivity is comparable to other methods. There is also a certain difference between the methods

Method used

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  • Pesticide ethofenprox fluorescence quantitative immunochromatography test paper and application thereof
  • Pesticide ethofenprox fluorescence quantitative immunochromatography test paper and application thereof
  • Pesticide ethofenprox fluorescence quantitative immunochromatography test paper and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1. Antigen synthesis and antibody preparation of the pesticide etofenprox

[0069] 1. Materials and methods

[0070] 1. Main reagents, materials and experimental animals

[0071] Succinic anhydride, 3-benzyloxybenzyl alcohol, bovine serum albumin (BSA), chicken ovalbumin (OVA), N-hydroxysuccinimide (NHS), N,N-dimethylformamide (DMF ), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), and p-aminophenylacetic acid were purchased from Sigma Company in the United States; sodium nitrite, hydrochloric acid, and sodium hydroxide were purchased from From Beijing Sinopharm Group; Freund's complete adjuvant and Freund's incomplete adjuvant were purchased from Gibco; Etoproxil standard was purchased from Beijing Century Aoke Biotechnology Co., Ltd.; tetramethylbenzidine (TMB), biotinylated Goat anti-rabbit IgG and avidinized HRP were purchased from Dakowei Biotechnology Co., Ltd.; Protein G antibody purification prepacked column (1mL) was GE products; EL...

Embodiment 2

[0115] Example 2, Preparation and application of the fluorescent quantitative immunochromatography test paper for the pesticide etofenproxil

[0116] 1. Materials and methods

[0117] 1. Experimental materials

[0118] M-OVA prepared in Example 1, rabbit anti-M-BSA polyclonal antibody (referred to as Ab-M, i.e. antibody 2 in Table 1 of Example 1); etofenprox, bifenthrin, permethrin standard Purchased from Beijing Century Aoke Biotechnology Co., Ltd.; other chemical reagents were purchased from Beijing Wangjia Biotechnology Co., Ltd.; fluorescent microspheres (diameter 330nm, excitation light 475nm, fluorescence 525nm, OCEANNANO TECH, product number 110315), nitrocellulose ( NC) film, sample pad (glass fiber material), marker pad (non-woven material), water-absorbing pad (paper), polyvinyl chloride (PVC) backboard, and test paper card plastic casing were all purchased from Beijing Jinyuanjiahe Biotechnology Co., Ltd.; IsoFlow Reagent Dispenser and Guillotine Cutter are produc...

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Abstract

The invention discloses pesticide ethofenprox fluorescence quantitative immunochromatography test paper and application thereof. The invention provides a fluorescent quantitative immunochromatographytest strip for pesticide ethofenprox, which comprises: a sample pad and a marker combination pad (with an anti-ethofenprox antibody marked by fluorescent microspheres) which are sequentially connectedand fixed on a bottom plate; the chromatographic membrane that is provided with a detection line (close to the sample pad and coated with a conjugate of phenylacetic acid-ethofenprox azo compound andcarrier protein 1) and a quality control line (close to the water absorption pad and coated with an antibody for resisting the ethofenprox antibody marked by the fluorescent microspheres); and a water absorption pad. When the test paper provided by the invention is used for detecting the pesticide ethofenprox, the detection limit is low, the accuracy is high, the specificity is strong, the precision is good, the standard curve equation R2 value can reach 0.9801, and a detection result can be obtained within 10min. Therefore, the prepared ethofenprox fluorescence quantitative immunochromatographic test paper card is suitable for rapid and quantitative detection of ethofenprox pesticides.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fluorescent quantitative immunochromatographic detection test paper for the pesticide etofenprox and its application. Background technique [0002] Pesticide residues are increasingly becoming a concern in agriculture and medicinal plant production, food and drug safety, and import and export industries. Especially in the export trade of Chinese medicinal materials, excessive pesticide residues often become a "stuck neck" factor, causing great losses to our country. Recently, the relevant state departments have increasingly strict requirements on pesticide residues, so the detection of pesticide residues has also attracted increasing attention. [0003] At present, the commonly used methods for detecting pesticide residues are mainly divided into two categories: one is to use large-scale or precision instruments for analysis, such as high-performance liquid chromatography (HPLC), ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/543G01N33/533C07K16/44C07K16/06
CPCG01N33/558G01N33/533G01N33/54313C07K16/44C07K2317/10G01N33/184
Inventor 吕俊海杨静修成奎
Owner EXPERIMENTAL RES CENT CHINA ACAD OF CHINESE MEDICAL SCI
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