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

A technology for immunofluorescence detection and ciprofloxacin, which is used in fluorescence/phosphorescence, measurement devices, preparation of samples for testing, etc.

Inactive Publication Date: 2013-10-30
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

Examples

Experimental program
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Effect test

Embodiment 1

[0048] Example 1. Composition and preparation of detection kit

[0049] Ciprofloxacin was purchased from Beijing Hengyuan Qitian Institute of Chemical Technology, and 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, the molecule itself has a carboxyl group, which can be directly coupled with protein.

[0050] 1. The original ciprofloxacin coating

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

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

[0053] 1) Take 33.1mg of ciprofloxacin, 3ml of N,N-dimethylformamide (DMF) and 3ml of dioxane and mix to obtain mixed solution a. The mixing temperature is 25℃ and the mixing time is 30min. The way is shock;

[0054] 2) 6ml of the m...

Embodiment 2

[0074] Example 2. Preparation method of standard curve

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

[0076] The detection of competitive detection is limited to the mass concentration of the competitive drug when B0 / B=1.2 (B0 is the detection value of the 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 test results are shown in Table 1...

Embodiment 3

[0080] Example 3. Determination of cross-reaction

[0081] Choose Enrofloxacin (Beijing Hengyuan Qitian Chemical Technology Research Institute, 30235CDCT-C13170000), Danofloxacin (Sigma-aldrich, 33700-100MG-R), Norfloxacin (Beijing Hengyuan Qitian Chemical Technology Research Institute, 30234CDCT-C15648000), Enoxacin (Beijing Hengyuan Qitian Institute of Chemical Technology, 29641NIC-130453), Ofloxacin (Beijing Hengyuan Qitian Institute of Chemical Technology, 30237CDCT-C15717000) and Penoxa Stars (Sigma-aldrich, 33899-100MG-R) were prepared into a series of concentrations and tested with a quantum dot kit. Calculate the IC50 of each competitor, and use the following formula to calculate the cross-reaction rate of these 5 drugs and the ciprofloxacin quantum dot kit. The calculation formula is: cross-reaction rate (%)=[IC50(ciprofloxacin) / IC50(drug under test)]×100.

[0082] The measurement and calculation results are shown in Table 2. The results showed that the ciprofloxacin qu...

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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 quantum dot-based immunofluorescence detection method for ciprofloxacin and a special kit. Background technique [0002] Quinolones (QNS) drugs are a very important class of broad-spectrum antibiotics that have developed rapidly in the past 20 years. They can inhibit bacterial DNA helicase, have 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. It is widely used in treatment, prevention and growth promotion. It has attracted widespread attention due to its drug resistance and potential carcinogenicity. In the tissues, the labeled residues of ciprofloxacin are ciprofloxacin and ciprofloxacin, of which the concentration of residues in liver tissue and kidney tissue is the highest, followed by skin tissue attached to muscle and fat, ciprofloxacin Ciprofloxacin (CIP), a metabolite of Star, stil...

Claims

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Application Information

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Patent Type & Authority Patents(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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