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Method for simultaneously detecting residual amount of quinolone antibiotics

A technology for quinolones and antibiotics, applied in the field of simultaneous detection of quinolone antibiotics residues, can solve the problems of complex technical operations, expensive equipment, high detection limits, etc., and achieve the effects of improving detection efficiency, shortening detection time, and simple experimental operations

Inactive Publication Date: 2018-08-28
NANJING XIANGZHONG BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The advantages of microbiological method are simple operation, low cost and rapid detection, but the detection limit of this method is higher than the minimum detection limit stipulated by various countries; the sensitivity is not high, and the specificity is poor
Although the detection results of chromatography are accurate, there are disadvantages such as complicated technical operation, long detection time, cumbersome pretreatment, expensive equipment, and high cost, so it is not suitable for large-scale on-site rapid detection
As a convenient, low-cost, fast and high-throughput screening method, enzyme-linked immunosorbent (ELISA) assay and colloidal gold test strip technology are widely used in many fields, but ELISA can only detect single-component residues. , while colloidal gold test strip technology is prone to high false positive or false negative rates

Method used

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  • Method for simultaneously detecting residual amount of quinolone antibiotics
  • Method for simultaneously detecting residual amount of quinolone antibiotics
  • Method for simultaneously detecting residual amount of quinolone antibiotics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A method for simultaneously detecting 9 kinds of quinolone antibiotic residues:

[0037] (1) Preparation of visualized protein chip: prepare a 6×5 matrix on a 96-well plate with a biochip spotter, and spot nine quinolones (ofloxacin, lomefloxacin, pefloxacin, Enoxacin, norfloxacin, enrofloxacin, ciprofloxacin, nalidixic acid, flumequine) monoclonal antibody, the sample volume is 50nL / point, and each sample is repeated 3 points; After the end, incubate in a 37°C incubator for 2 hours to immobilize the antibody on the bottom of the plate; add 200 μL chip blocking agent (BSA) to each well, incubate in a 37°C incubator for 2 hours, and then wash with washing buffer 3 times. Each time for 10 seconds, pat dry, and store in a 4°C refrigerator for later use;

[0038] (2) Add 25 μL (ofloxacin, lomefloxacin, pefloxacin, enoxacin, norfloxacin, enrofloxacin, cyclofloxacin, Profloxacin, nalidixic acid, flumequine) mixed standards or samples with different concentration gradients, ...

Embodiment 2

[0049] Specificity test for quinolones

[0050] The specificity of quinolone antibodies is usually judged using competition inhibition curves. Different concentrations of antigen and interferent were used to calculate their respective binding ratios (B / B 0 ), draw competitive inhibition curves, and calculate their respective ICs 50 . The cross-reactivity rate was calculated according to the following formula (1).

[0051] CR=[IC 50 (analyte) / IC 50 (interference object)]×100% (1)

[0052] IC 50 is the concentration required for 50% inhibition of the analyte or interferent-induced signal.

[0053] The experimental results are shown in Table 2, indicating that the cross-reactivity rates between quinolone antibodies and nine quinolone drugs are all between 1% and 5%. It shows that the specificity of the quinolone antibody is good, there is no crossover, and each quinolone antibiotic can be detected separately.

[0054] Table 2 Quinolones cross-reaction rate

[0055]

...

Embodiment 3

[0058] Establishment of standard curve for quinolones

[0059] The indirect competition method was used to explore the standard curve of quinolones. 3 parallel microarray points for each antibody concentration point, the signal value is taken as the average value, within a certain range, with the increase of the concentration of the competition standard, the content of the labeled artificial antigen bound by the fixed antibody will be less and less, Therefore, the detected signal value will be lower. After exceeding this range, the detected signal value will not change with the increase of the concentration of the competition standard. Use standard products of ofloxacin, standard products of lomefloxacin, standard products of pefloxacin, standard products of enoxacin, standard products of norfloxacin, standard products of enrofloxacin, standard products of ciprofloxacin, naphthalene Glycolic acid standard substance, flumequine standard substance (as shown in table 3) mixed st...

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Abstract

The invention discloses a method for simultaneously detecting residual amount of quinolone antibiotics. By adopting a direct competition law reaction principle, monoclonal antibodies of a plurality ofquinolone medicines are fixed to the bottom of a microporous plate in a micro-array form so as to prepare a microporous plate biological chip array; reaction on the microporous plate biological chiparray as well as hybrid artificial antigen marked with a quinolone nano-material and a quinolone medicine standard product is sequentially carried out; finally, color development is increased by usingsilver; moreover, a microporous color development result is quickly scanned and imaged by using a visualized chip scanning instrument, and images are processed by adopting chip analysis software. According to the method disclosed by the invention, the detection time is greatly shortened, the flexibility is improved, and an experiment is easy to operate; nine antibiotics of the quinolones can be simultaneously and separately detected, so that the detection efficiency is improved; moreover, significance on researching a method for analyzing multiple medicine residues of the quinolones is realized, the method disclosed by the invention has good application and promotional value, and provides a certain reference for detecting the residues of the quinolones in the food.

Description

technical field [0001] The invention belongs to the detection technology of drug residues, in particular to a method for simultaneously detecting the residues of quinolone antibiotics. Background technique [0002] Quinolones (Fluoroquinolones, FQs) are a class of antibacterial drugs with a 6-fluoro-7-piperazine-4-norone ring structure (mother core structure). Quinolones are a class of broad-spectrum antibacterial drugs developed in recent years, and they are also new varieties that have been developed and applied by countries all over the world in recent years. So far, many drugs have been put on the market. In order to control and treat animal diseases, multiple quinolones are used simultaneously or alternately in the process of breeding and treating animals, which leads to the occurrence of multiple drug residues. Exceeding the standard of quinolone drug residues in animal food is a potential threat to the human body, and the problem of veterinary drug residues has attra...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 李周敏许丹科李钟卉姜金斗
Owner NANJING XIANGZHONG BIOTECH
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