Near-infrared fluorescent microsphere immunochromatography test strip rapid detection method

A technology of fluorescent microspheres and detection methods, which is applied in the direction of fluorescence/phosphorescence, material analysis through optical means, and measurement devices, which can solve the problems of low detection sensitivity and inability to detect multiple antibiotics, and achieve the effect of shortening the detection time

Inactive Publication Date: 2021-08-24
WWHS BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the detection sensitivity of the immunochromatographic test strips reported so far is low, and most of them are mainly used to detect a single antibiotic, and cannot detect multiple antibiotics at the same time.

Method used

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  • Near-infrared fluorescent microsphere immunochromatography test strip rapid detection method
  • Near-infrared fluorescent microsphere immunochromatography test strip rapid detection method
  • Near-infrared fluorescent microsphere immunochromatography test strip rapid detection method

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preparation example Construction

[0073] In a preferred embodiment, the preparation method of the analysis test strip comprises the following steps:

[0074] Step 1, prepare the sample pad: the sample pad is made of glass fiber, cut the sample pad into a strip of 30cm×28mm, spread it on the grid plate, take 7mL containing 1.2% Tris, 0.4% BSA, 0.4% Tween -20. Treat the sample pad with 0.12% EDTA phosphate buffer solution to make it fully wet, then place it in an electric blast drying oven at 37°C for 12 hours until it is dried;

[0075] Step 2: Treat the nitrocellulose membrane and dilute the sulfonamide antigen, quinolone antigen, lincomycin antigen and goat anti-rabbit secondary antibody to the desired concentration with a coating solution containing 2% trehalose and 5% methanol , spray the above four solutions onto the NC membrane at a speed of 0.8 μL / cm in turn with a film-drawing device, and use them as the detection line (T1 line, T2 line, T3 line) and quality control line (C line) of the test strip respe...

Embodiment 1

[0084] Near-infrared fluorescent microspheres are monodisperse polystyrene microspheres coated with a large number of near-infrared II region fluorophores and rich in functionalized carboxyl groups on the surface. The characterization method includes the following steps:

[0085] (1) The near-infrared fluorescent microspheres were stored in ultrapure water in the form of a 10 mg / mL dispersion. In order to characterize its microscopic morphology, a scanning electron microscope was selected to observe its surface morphology. The fluorescent microsphere powder was fixed on the electron microscope stage with a double-layer conductive adhesive, and the floating particles that were not adhered were blown off with an ear cleaning ball. SEM used 3.0 Accelerating voltage of kV is magnified by 70.0k times for scanning imaging. The surface morphology of near-infrared fluorescent microspheres is as follows: figure 2 shown.

[0086] (2) Select the transmission electron microscope to obs...

Embodiment 2

[0089] The near-infrared fluorescent microsphere immunochromatographic test strip detection method of common antibiotics in milk comprises the following steps:

[0090] (1) Preparation of near-infrared fluorescent microsphere probes: use the initial washing solution to disperse and wash the near-infrared fluorescent microspheres three times; add 25 μL of EDC with a concentration of 10 mg / mL and 75 μL of EDC with a concentration of 10 mg / mL to the above solution The sulfo-NHS aqueous solution was activated at room temperature for 10 min in a rotating state; the above-mentioned activated fluorescent microspheres were washed with a coupling solution, and then an appropriate amount of antibody was added, and coupled at room temperature for 3 h in a rotating state, during which ultrasonic dispersion was performed several times to avoid aggregation of the microspheres; Add 30 μL of 10% ethanolamine aqueous solution to the above-mentioned fluorescent microspheres after coupling, and b...

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Abstract

The invention provides a near-infrared fluorescent microsphere immunochromatography test strip rapid detection method for common antibiotics in milk, and the method comprises the following steps: S1, preparing a PVC base plate, and a 2mm sample pad, a nitrocellulose membrane and absorbent paper which are overlapped and pasted on the base plate; treating the sample pad through a sample pad diluent, and coating the nitrocellulose membrane with an artificial antigen of antibiotics as a detection line and coated with a goat anti-rabbit antibody as a quality control line; and S2, respectively coupling the near-infrared fluorescent microspheres with a sulfanilamide monoclonal antibody, a quinolone monoclonal antibody, a lincomycin monoclonal antibody and a rabbit anti-goat antibody. According to the application, an immunochromatography test strip system can be effectively improved; the detection method has the advantages of high sensitivity, good stability and low cost, and can be used for on-site rapid determination of sulfanilamide, quinolone and lincomycin in milk.

Description

[technical field] [0001] The invention relates to the technical field of food safety immunological detection, and relates to a rapid detection method for near-infrared fluorescent microsphere immunochromatographic test strips with many residues of sulfonamides, quinolones and lincomycin. [Background technique] [0002] Sulfonamide is a general term for a class of components with p-aminobenzenesulfonamide as the basic structure. Sulfonamide drugs are their synthetic derivatives. They are the first type of artificially synthesized antibacterial drugs and are widely used in animal husbandry. There are more than 20 kinds of sulfa drugs commonly detected in food. The antibacterial effect of sulfonamides was discovered as early as the 1940s, and it has become an effective treatment method for bacterial infections in livestock and poultry. It has a bacteriostatic effect and has a good killing effect on Gram-positive bacteria, some Gram-negative bacteria and a variety of coccidiosi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N21/64
CPCG01N33/558G01N21/643
Inventor 王国新张昊廖滔
Owner WWHS BIOTECH INC
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