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Method and special ELISA reagent kit for detecting nitryl imidazole medicament

An enzyme-linked immunosorbent reagent, nitroimidazole technology, applied in immunoglobulin, chemical instruments and methods, cells modified by introducing foreign genetic material, etc., can solve the problems of high cost, limited popularization and application, cumbersome and complicated operation, etc. , to achieve the effect of low cost, simple processing process and high sensitivity

Inactive Publication Date: 2008-12-03
BEIJING WANGER BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods for the determination of nitroimidazoles mainly include high performance liquid chromatography, liquid chromatography tandem mass spectrometry, etc., which are complicated and expensive to operate, and their popularization and application are limited.

Method used

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  • Method and special ELISA reagent kit for detecting nitryl imidazole medicament
  • Method and special ELISA reagent kit for detecting nitryl imidazole medicament
  • Method and special ELISA reagent kit for detecting nitryl imidazole medicament

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Preparation and detection method of an enzyme-linked immunosorbent assay kit using a conjugate of a nitroimidazole hapten and a carrier protein as a coating source

[0035] 1. The enzyme-linked immunosorbent assay kit that uses the conjugate of nitroimidazole drug hapten and carrier protein as the coating source generally includes:

[0036] (1) A microtiter plate coated with a coating source (nitroimidazole hapten and carrier protein conjugate); the concentration of the coating source can be 0.05-0.1 μg / ml.

[0037] (2) Horseradish peroxidase-labeled goat anti-mouse anti-antibody working solution: the diluent of enzyme-labeled secondary antibody is phosphate buffer containing 0.5% bovine serum albumin, and the dilution concentration of goat anti-mouse anti-antibody is 1:1000 ; The percentage is a mass percentage.

[0038] (3) Metronidazole standard solution (purchased from Dr. Germany, product number: RBDR-C15201000): a total of 6 bottles, the concentrations...

Embodiment 2

[0101] Example 2. Preparation and detection method of ELISA kit with anti-antibody as coating source

[0102] 1. ELISA kits with anti-antibodies as the coating source generally include:

[0103] (1) ELISA plates coated with anti-antibodies; the concentration of anti-antibodies is 0.05-0.1 μg / ml;

[0104] (2) The working solution concentration of horseradish peroxidase-labeled nitroimidazole drug antigen is 0.15M; it is prepared according to the following method: the horseradish peroxidase-labeled ronidazole half Dilute the antigen to a concentration of 0.15M, 12ml / bottle, 1 bottle.

[0105] (3) Metronidazole standard solution (purchased from Dr. Germany, product number: RBDR-C15201000) solution: a total of 6 bottles, the concentrations were 0 μg / L, 0.1 μg / L, 0.3 μg / L, 0.9 μg / L, 2.7 μg / L, 8.1μg / L. The diluent is pH7.4, 0.05M phosphate buffer.

[0106] (4) Substrate chromogenic solution: The substrate chromogenic solution is composed of A liquid and B liquid, the substrate c...

Embodiment 3

[0140] Embodiment 3, test kit sensitivity, accuracy and shelf life test

[0141] (1) Kit sensitivity experiment

[0142] Get the negative chicken sample that does not contain nitroimidazoles medicine and carry out 20 times of detection respectively with the prepared kit of the present invention, the mean value of measurement result plus 3 times of standard deviation is as the minimum detection limit of kit.

[0143] Table 1-1 Statistical Table of Determination Results of Negative Chicken Samples μg / kg

[0144] (The original coating is a conjugate of nitroimidazole hapten and carrier protein)

[0145]

[0146] Table 1-2 Statistical Table of Determination Results of Negative Chicken Samples μg / kg

[0147] (Coating was originally anti-antibody)

[0148]

[0149] It can be seen from Table 1-1 and 1-2 that the minimum detection limit of the kit is 0.1 μg / kg.

[0150] (2) Precision test of standard products:

[0151] 10 test kits were extracted from each batch of the three ...

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Abstract

The invention discloses a method for detecting nitroimidazole drugs and a dedicated enzyme-linked immune kit. The kit comprises a special antibody, and a coating antigen and an enzyme label; nitroimidazole is at least one of the following components: metronidazole, tinidazole, imetridazole and ronidazol. The method in the invention has the advantages of simple sample pretreatment process, simple operation, low cost, good specificity, high sensitivity, high accuracy, etc.; the method can perform on-site monitoring and is suitable for screening a plurality of samples. Therefore, the method and the dedicated kit in the invention play an important part in detecting the residues of the nitroimidazole drugs in animal derived food.

Description

technical field [0001] The invention relates to a method for detecting nitroimidazole drugs and a special ELISA kit. Background technique [0002] Nitroimidazoles include metronidazole, metronidazole, clonidazole, ronidazole and dimetidazole, etc., and their general chemical structures are as follows: figure 1 Shown (in the figure, when R1 and R2 are both methyl, the drug is dimetidazole, when R1 is H, and R2 is ethanol, the drug is metronidazole). This type of drug is often used to prevent and treat specific bacteria and protozoan diseases. The principle is that this type of drug inhibits the redox reaction of the protozoan, breaks the nitrogen chain of the pathogen, and has obvious effects of killing amoeba and anti-anaerobic bacteria in vitro. It is effective on all parts of the amoeba, rapidly absorbed after oral administration, and widely distributed in various organs and most body fluids in the body. However, because of its potential carcinogenicity and mutagenicity,...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577G01N33/535C12N5/12C07D233/00C07K16/00
Inventor 沈建忠何方洋丁双阳冯才伟万宇平冯静汪善良朱亮罗晓琴
Owner BEIJING WANGER BIOTECH
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