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Quantum dot-based method for detecting ciprofloxacin by immunofluorescence and special kit

A technology for immunofluorescence detection and ciprofloxacin, which is applied in the direction of fluorescence/phosphorescence, measurement devices, and preparation of test samples

Inactive Publication Date: 2011-11-23
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Since the quantum dot labeling materials suitable for immunodiagnostic reagents must meet the requirements of high quantum yield, strong fluorescence, good biocompatibility, high stability and low cost, the quantum yield of the current foreign commercialized quantum dots is mostly in the range of 40 % or less, its size is relatively small, and it needs to be irradiated and excited by laser optical instruments. At present, it can only be used in cell immunofluorescence detection and flow cytometry detection and sorting. Therefore, there is no quantum dot immunoassay reagent on the international market.

Method used

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  • Quantum dot-based method for detecting ciprofloxacin by immunofluorescence and special kit
  • Quantum dot-based method for detecting ciprofloxacin by immunofluorescence and special kit
  • Quantum dot-based method for detecting ciprofloxacin by immunofluorescence and special kit

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Effect test

Embodiment 1

[0048] Embodiment 1, composition and preparation of detection kit

[0049] Ciprofloxacin was purchased from Beijing Hengyuan Qitian Chemical Technology Research Institute, the product catalog number is 30233CDCT-C11668500. Its chemical name is 1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-3-quinolinecarboxylic acid. Ciprofloxacin is a quinolone drug, and the molecule itself has a carboxyl group, which can be directly coupled with proteins.

[0050] 1. Ciprofloxacin coated original

[0051] The conjugate of ciprofloxacin and carrier protein BSA, the ciprofloxacin and BSA are connected through the peptide bond formed by the carboxyl group in ciprofloxacin and the amino group in BSA.

[0052] The preparation method of ciprofloxacin coating former is as follows:

[0053] 1) Mix 33.1mg of ciprofloxacin, 3ml of N,N-dimethylformamide (DMF) and 3ml of dioxane to obtain a mixed liquid a. The mixing temperature is 25°C and the mixing time is 30min. The method is shock; ...

Embodiment 2

[0074] Embodiment 2, the preparation method of standard curve

[0075] Add the ciprofloxacin standard solution that concentration is 0,0.001,0.005,0.01,0.05,0.1,0.5,1,5,10ug / L in the prepared ciprofloxacin microwell strip, 50ul / hole, will Dilute quantum dot-labeled CIP antibody with PBS-T diluent 1:50 [25ug labeled antibody: 5ml diluent], add 50ul to each microwell, shake at room temperature for 1 hour, wash with washing solution 3 times and use a fluorescent microplate reader Check its fluorescence intensity value. The fluorescent microplate reader was set to an excitation wavelength of 345 nm and an emission wavelength of 620 nm.

[0076] The detection of the competition detection is limited to the mass concentration of the competitive drug when B0 / B=1.2 (B0 is the detection value of the 0 standard sample, B is the detection value of the test sample), and the sensitivity of the detection system is determined according to the curve regression equation. The detection result...

Embodiment 3

[0080] Embodiment 3, the mensuration of cross reaction

[0081] Choose enrofloxacin (Beijing Hengyuan Qitian Chemical Technology Research Institute, 30235CDCT-C13170000), dafloxacin (Sigma-aldrich, 33700-100MG-R), norfloxacin (Beijing Hengyuan Qitian Chemical Technology Research Institute, 30235CDCT-C13170000), Institute, 30234CDCT-C15648000), enoxacin (Beijing Hengyuan Qitian Chemical Technology Research Institute, 29641NIC-130453), ofloxacin (Beijing Hengyuan Qitian Chemical Technology Research Institute, 30237CDCT-C15717000) and Penoxa Star (Sigma-aldrich, 33899-100MG-R) was formulated into a series of concentrations, and detected with a quantum dot kit. Calculate the IC50 of each competitor, and use the following formula to calculate the cross-reactivity rate of these 5 kinds of drugs and the ciprofloxacin quantum dot kit respectively. The calculation formula is: cross-reaction rate (%)=[IC50 (ciprofloxacin) / IC50 (drug to be tested)]×100.

[0082] The measurement and cal...

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Abstract

The invention discloses a quantum dot-based method for detecting ciprofloxacin by immunofluorescence and a special kit. The immunofluorescence detection kit for detecting the ciprofloxacin comprises a ciprofloxacin peridium and a ciprofloxacin antibody marked by a quantum dot. By the method, the quantitative detection of ciprofloxacin residual medicines can be realized, the detection limit is low, the detection sensitivity is high, the specificity is high, and the method is suitable for detecting various samples. Therefore, the method has a bright application prospect in the field of the detection of the ciprofloxacin.

Description

technical field [0001] The invention relates to a method for detecting ciprofloxacin by immunofluorescence based on quantum dots and a special kit. Background technique [0002] Quinolones (QNS) drugs are a class of very important broad-spectrum antibiotics that have been rapidly developed in the past 20 years. They can inhibit bacterial DNA helicase, have a broad 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 has been widely used in treatment, prevention and growth promotion, and has attracted widespread attention due to its drug resistance and potential carcinogenicity. In tissues, the marked residues of ciprofloxacin are ciprofloxacin and ciprofloxacin, among which the residue concentration in liver tissue and kidney tissue is the highest, followed by muscle and fat-attached skin tissue, ciprofloxacin Star its metabolite ciproflox...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/532G01N33/544G01N1/38G01N21/64
Inventor 马岚袁航吴峰
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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