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Chloramphenicol quantitative detection method based on up-conversion phosphor technology and immunochromatography technology

A technology of immunochromatography and chloramphenicol, which is applied in the fields of nano-biomarkers, optical detection, and immunology, can solve the problems of inability to apply sample on-site detection or accurate quantification, inability to use on-site detection, unreliable results, etc., to avoid this problem Low interference, saving steps, and reducing false positive effects

Inactive Publication Date: 2015-02-25
BEIJING UNIV OF TECH
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Problems solved by technology

The biggest advantage of immunochromatography technology is that the detection time is fast. Generally, the detection time is 10-15 minutes. It does not require large-scale equipment and can be used for real-time detection. Generally, colloidal gold is the most common marker and can be used for various foods, agricultural products, diseases, etc. The detection time is fast, but there are two disadvantages: one is that the colloidal gold marker is prepared by physical adsorption, and it is easy to dissociate during the chromatography process, resulting in false positives
The second is to judge the result by color reaction, the result is unreliable, and the sensitivity is low.
Chromatography can significantly improve the detection sensitivity, but the operation is complicated and the steps are cumbersome, so it cannot be used for on-site detection; although the detection time of colloidal gold immunochromatography is shortened, the disadvantages of high false positive and low sensitivity limit its application; Repeated washing, long detection time
In short, these detection techniques are time-consuming and labor-intensive, and the detection cost is high, so they cannot be applied to the on-site detection of samples or the purpose of accurate quantification
[0005] The invention discloses a method for quantitative detection of chloramphenicol based on up-conversion luminescence technology and immunochromatography technology. After literature research, it is found that the combination of up-conversion luminescence technology and immunochromatography technology for chloramphenicol has not been reported at home and abroad. Examples of element detection

Method used

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  • Chloramphenicol quantitative detection method based on up-conversion phosphor technology and immunochromatography technology
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  • Chloramphenicol quantitative detection method based on up-conversion phosphor technology and immunochromatography technology

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Embodiment 1

[0033] Upconversion nanomaterial used: NaYF 4 :Yb 3+ ,Er 3+

[0034] (1) Add the surfactant polyoxyethylene nonylphenyl ether (IGEPALCO520) into 145ml cyclohexane, ultrasonically disperse it into a transparent homogeneous solution with a concentration of 0.05g / ml, weigh up-conversion nanoparticles and add them to the cyclohexane Hexane, with constant ultrasonic stirring to form a transparent solution with a concentration of upconverting nanoparticles of 0.013%. Measure 0.58ml of an aqueous solution of 28% ammonia gas and add it to the above reaction system. When a little milky white colloid appears in the solution, ultrasonically and continuously stir to form a transparent homogeneous solution again, and finally add 0.145ml of tetraethyl orthosilicate. The reactor was placed on a magnetic stirrer and stirred at room temperature and pressure for 16 h. When the solution turned from transparent to light white, add a little methanol to precipitate the particles until saturated...

Embodiment 2

[0043] Others are the same as Example 1, and (4) and (7) are prepared as follows:

[0044] (4) Prepare a chromatographic solution containing 0.1% Tween-20 in volume fraction, 0.1% BSA and 1% sucrose in mass concentration, use 0.03mol / l phosphate buffer as the solvent, adjust the pH to 7.2, and take out the upconversion The suspension of the marker was centrifuged to remove the supernatant, and the precipitate was resuspended with the above chromatography solution for later use;

[0045] (7) Dilute 1mg / ml chloramphenicol by 100-100000 times with the above-mentioned chromatographic solution into standard solutions of different concentration gradients, use the chromatographic solution that does not contain chloramphenicol as a negative control, get each concentration standard solution and Control solution 0.1ml is added to 20ul mass concentration and is the suspension of the up-conversion marker of 1mg / ml and mixes evenly, the sample pad of the test paper prepared in embodiment 1 (...

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Abstract

The invention relates to a chloramphenicol quantitative detection method based on an up-conversion phosphor technology and an immunochromatography technology, belonging to the fields of nano biomarkers, immunology and optical detection. The detection method mainly comprises the following steps that carboxylation modification is carried out on 200-300nm up-conversion fluorescent nanoparticles so that the up-conversion fluorescent nanoparticles become markers which are good in water solubility and high in dispersibility and can be easily coupled with biomolecules; a sample pad, a conjugate pad treated by the up-conversion markers, a nitrocellulose membrane (NC membrane) treated by antigen-antibody sample application and absorbent paper are combined together by a sticky bottom lining; antigen takes chloramphenicol-bull serum albumin (BSA) as a detection line (T), antibody takes goat-anti-mouse antibody as a quality control line (C), and the distance between the detection line and the quality control line is 0.5cm; the assembled test paper is cut into test paper strips which are 6cm long and 4mm wide by a strip cutting machine, the test paper strips are assembled into a shell to establish the immune chromatography test paper; chloramphenicol standard antigens with different concentration gradients are loaded to the sample pad in the test paper shell; after standing still for 10-15 minutes, the sample pad is put into an up-conversion detector for detection. The chloramphenicol quantitative detection method is simple in preparation technology and operation, can be completed without a complicated instrument or equipment, is rapid and sensitive, and has an important significance for accurately and quantitatively testing the content of chloramphenicol in food.

Description

technical field [0001] The present invention combines up-conversion luminescence technology and immunochromatography technology, and is characterized in that the up-conversion marker is used as a detection molecule, a test paper is used as a solid phase carrier, and a laser detector is used as a detection means. The new detection technology belongs to the field of nano-biological markers, The field of immunology and the field of optical detection. Background technique [0002] Up-conversion luminescence technology is a fluorescence detection technology that has emerged in recent years. Up-conversion fluorescent materials are generally composed of single rare earth-doped fluoride or oxide and double-doped fluoride. The principle is that under the excitation of near-infrared light, Emits visible fluorescence. From a microscopic point of view, the absorber in the material mainly absorbs infrared photons and transitions from the ground state to the excited state, and then the a...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N21/64
CPCG01N33/558G01N21/64
Inventor 马雪梅於然谢飞
Owner BEIJING UNIV OF TECH
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