Surface-enhanced Raman gold-labeled test paper strip for quickly detecting trace fipronil and preparation method thereof

A surface-enhanced Raman and micro-fipronil technology, which is applied in the direction of measuring devices, Raman scattering, instruments, etc., to achieve high sensitivity, consumer health protection, and high labeling rate

Inactive Publication Date: 2018-04-20
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surface-enhanced Raman immunoassay has the characteristics of high sensitivity of Raman, and also has the advantage of strong antibody specificity. It has unique advantages for the screening of fipronil. Ho

Method used

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  • Surface-enhanced Raman gold-labeled test paper strip for quickly detecting trace fipronil and preparation method thereof
  • Surface-enhanced Raman gold-labeled test paper strip for quickly detecting trace fipronil and preparation method thereof
  • Surface-enhanced Raman gold-labeled test paper strip for quickly detecting trace fipronil and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] like Figure 1-3 It is given that the surface-enhanced Raman gold standard test strip for rapid detection of trace amounts of fipronil in the present invention, the bottom layer is a support layer, the middle layer is an adsorption layer, the protective layer is fixed on the adsorption layer, and the adsorption layer is sequentially from the test end to the adsorption fiber Layer, surface-enhanced Raman colloidal gold-labeled antibody fiber layer, cellulose film layer and water-absorbing material layer at the handle end, wherein the carrier protein coupled to fipronil is sprayed on the cellulose film as a detection line, and there is also a sheep antibody or Rabbit anti-mouse IgG, goat anti-rabbit IgG antibody or SPA printed quality control lines.

[0037] The labeled antibody in the surface-enhanced Raman colloidal gold-labeled antibody fiber layer is a surface-enhanced Raman colloidal gold-labeled anti-fipronil monoclonal antibody or polyclonal antibody.

[0038] The...

Embodiment 2

[0044] In order to make rapid detection trace fipronil surface-enhanced Raman colloidal gold test strips, first prepare the coupling fipronil carrier protein and fipronil surface-enhanced Raman colloidal gold-labeled antibody, so as to prepare the detection line and surface-enhanced Raman Colloidal gold labeling antibody fiber layer; secondly prepare goat anti or rabbit anti-mouse IgG (or goat anti-rabbit IgG) antibodies for the preparation of quality control lines.

[0045] The preparation method of the surface-enhanced Raman gold standard test strip for rapid detection of trace amounts of fipronil of the present invention comprises the following steps:

[0046] 1. Preparation of carrier protein coupled to fipronil (i.e. fipronil carrier protein conjugate)

[0047] (1) The hydrolysis method is used to modify the structure of fipronil. Under the condition of strong alkali, the cyano group (-CN) in fipronil (CNT) is hydrolyzed into carboxyl group (-COOH) to obtain CNT-COOH. Di...

experiment example 1

[0063] Experimental Example 1, Sensitivity Detection

[0064] 1. Visual inspection with naked eyes:

[0065] Using fipronil solutions with concentrations of 1ng / mL, 2ng / mL, 4ng / mL, 6ng / mL, 8ng / mL, 10ng / mL, 20ng / mL and 40ng / mL as samples, surface-enhanced Raman Gold standard test strips were used for the detection of fipronil. The results of visual inspection (5-10 minutes) showed that all the test strip test lines did not develop color at 10ng / mL, 20ng / mL and 40ng / mL, and 90% of the test strip test lines did not develop color at 8ng / mL At 6ng / mL, 60% of the test strip detection lines do not develop color, and the test strip detection lines of 1ng / mL, 2ng / mL and 4ng / mL all develop color, so it is determined that the fipronil surface-enhanced Raman gold standard The lowest visual detection concentration of the test paper is 10ng / mL, and the sensitivity is set at 10ng / mL.

[0066] 2. Raman instrument detection:

[0067] With concentrations of 4ng / mL, 2ng / mL, 1ng / mL, 0.5ng / mL,...

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Abstract

The invention discloses a surface-enhanced Raman gold-labeled test paper strip for quickly detecting trace fipronil and a preparation method thereof. A bottom layer of the test paper strip is a support layer, a middle layer of the test paper strip is an adsorption layer, a protection layer is fixed on the adsorption layer, and the adsorption layer sequentially comprises an absorption fibrous layer, a surface-enhanced Raman colloidal gold-labeled antibody fibrous layer, a cellulose membrane layer and a water absorption material layer of a handle end from a test end, wherein the cellulose membrane is coated with fipronil-coupled carrier protein to serve as a detection line and is also provided with a quality control line printed by goat or rabbit anti-mouse IgG (Immunoglobulin G), goat anti-rabbit IgG antibody or SPA (Staphylococcal Protein A). The test paper strip disclosed by the invention is prepared on the basis of a competitive mode, and target objects on a sample and the detectionline are competitively combined with the surface-enhanced Raman colloidal gold-labeled antibody. Tests show that the test paper has stronger specificity and higher sensitivity, the visual detection limit is an ng level, the quantitative detection limit of a Raman instrument is a pg level, and the test paper has no cross reaction with other pesticides or veterinary drugs.

Description

technical field [0001] The invention relates to a device for rapidly detecting trace amounts of pesticides, in particular to a surface-enhanced Raman gold standard test strip for rapidly detecting trace amounts of fipronil and a preparation method thereof. Background technique [0002] Fipronil, trade name Fipronil, English common name Fipronil (fipronil), chemical name (RS)-5-amino-1-(2,6-dichloro-a,a,a-trifluoro-p -Tolyl)-4-trifluoromethylsulfinylpyrazole-3-carbonitrile, the molecular formula is C 8 N 2 Cl 4 , molecular formula C12H4Cl2F6N4OS, molecular weight 437.2, is a phenylpyrazole insecticide developed in 1981 by the French company Rhône-Planck. It has been used since the mid-1990s. It is a broad-spectrum insecticide and veterinary drug widely used in the world. It is mainly used for crop pest control, family pet parasite treatment and control, public health and entertainment venue pest control, etc. Widely used in all aspects. It binds to the γ-aminobutyric aci...

Claims

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Application Information

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IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 胡骁飞邓瑞广魏凤仙王方雨杨艳艳孙亚宁姚静静邢广旭柴书军杨继飞邢云瑞
Owner HENAN ACAD OF AGRI SCI
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