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Fluorescent microsphere immunochromatography detection card for detecting enrofloxacin and preparation method thereof

A technology of fluorescent microspheres and enrofloxacin, which is applied in chemical instruments and methods, measuring devices, and analytical materials, etc., can solve problems such as inability to sample multiple dilutions, sensitivity limitations, and many interfering components, and achieve low prices, The effect of high sensitivity and increased fluorescence lifetime

Inactive Publication Date: 2011-04-20
江西中德生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, no matter what kind of principle is used for rapid reagents used in China, due to their sensitivity limitations, the sample cannot be diluted multiple times, so that there are many interfering components in the sample, resulting in false negatives or false positives in the actual sample detection process. Phenomenon

Method used

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  • Fluorescent microsphere immunochromatography detection card for detecting enrofloxacin and preparation method thereof
  • Fluorescent microsphere immunochromatography detection card for detecting enrofloxacin and preparation method thereof
  • Fluorescent microsphere immunochromatography detection card for detecting enrofloxacin and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. Preparation of fluorescent microsphere markers (EDC method):

[0044] Preparation of fluorescent microsphere-labeled anti-enrofloxacin antibody: Take 1 mg of fluorescent microspheres coated with isothiocyanatorhodamine organic dye and centrifuge at 1000×g for 10 minutes, collect the precipitate, and wash with 0.01M borate buffer solution of pH 4.8 Adjust the microsphere concentration to OD 450 =0.2, then add 90μl 50mg / ml EDC, 150μl 5mg / ml NHS, shake and mix well, incubate at room temperature for 20min, centrifuge at 1000×g for 5min, dissolve the precipitate with 0.01M borate buffer solution of pH4.8, and adjust micro Ball concentration is OD 450 is 0.5. at 0.1ml OD 450 Add 1 μg of anti-enrofloxacin antibody to the fluorescent microspheres with a pH of 0.5, mix well, stir and react at room temperature for 3 h, wash and centrifuge 3 times with ultrapure water, wash with 0.01M PBS of pH 7.2 (which contains 5% sucrose and 0.05% Tween-20) to resuspend the pellet to th...

Embodiment 2

[0056] The preparation method of the present embodiment is basically the same as that of Example 1, the difference is:

[0057] The carrier protein is ovalbumin, and the labeled carrier fluorescent microspheres are quantum dot-silica core-shell dual-structure microspheres.

[0058] The qualitative detection of enrofloxacin with the above-mentioned fluorescent microsphere immunochromatographic detection card includes the following steps:

[0059] (1) Lay the test card flat, after the chicken liver extract sample to be tested is equilibrated to room temperature, add 90 μl of sample into the sample hole of the test card, react at room temperature for 15 minutes, and put the test card into the detection window;

[0060] (2) The fluorescent microspheres trapped in the detection area and the quality control area emit strong fluorescence under the excitation of the best excitation light source; observe in the observation window, if there is a fluorescent band in the detection area of...

Embodiment 3

[0062] The preparation method of this example is basically the same as that of Example 1, the difference is: 1. The fluorescent microspheres of the labeling carrier are isothiocyanatofluorescein-silicon dioxide core double-structured microspheres, and the surface is modified by streptavidin and prime. 2. The anti-enrofloxacin antibody can be coupled with biotin by the EDC method, which is basically the same as in Example 1.

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Abstract

The invention discloses a fluorescent microsphere immunochromatography detection card for detecting enrofloxacin and a preparation method thereof. The detection card sequentially comprises filter paper, a sample pad, a glass fiber film, a nitrocellulose film and absorbent paper, wherein a fluorescent microsphere-labeled antibody is sprayed on the glass fiber film; a detection area and a quality control area are fixed on the nitrocellulose film; a conjugate of enrofloxacin and carrier protein is sprayed in the detection area; and an anti-mouse antibody is sprayed in the quality control area. By taking core-shell bistructure luminous nano particles complexed by silica and fluorescent substances as labels and adopting immunochromatography in a competition blocking mode, the invention realizes rapid immunoassay of the enrofloxacin. In the detection process, an optimized excitation light source of the fluorescent microsphere is adopted for excitation, the emitted fluorescence passes through a light filter device, and that whether a detection line is provided with fluorescent substances is observed by naked eyes. The invention has the characteristics that: the detection card has high sensitivity and is rapid in detection, convenient to operate, and economic and practical.

Description

technical field [0001] The invention belongs to the field of food safety detection, and in particular relates to a detection card for qualitative detection of enrofloxacin using fluorescent microsphere immunochromatography technology and a preparation method thereof. Background technique [0002] Enrofloxacin (ENR), also known as ethyl ciprofloxacin, is the first fluoroquinolone drug exclusively for livestock and poultry. It was first successfully developed by Bayer in Germany in 1983, and it was put on the market in my country in 1994. Because of its ability to inhibit bacterial DNA helicase, wide antibacterial spectrum, high efficiency, low toxicity, and strong tissue penetration, it has become one of the most important anti-infective drugs in veterinary clinics and aquaculture, and is widely used in the treatment and prevention of diseases and promote growth. The drug is rapidly absorbed by oral administration and intramuscular injection, and is metabolized into ciproflo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/533G01N33/566G01N33/52C09K11/06C09K11/08
Inventor 陈雪岚杨晓慧熊勇华赖卫华魏华李林
Owner 江西中德生物工程股份有限公司
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