Kit and method for detecting beta-lactam antibiotics

A lactam and kit technology, which is applied in the field of kits for detecting beta-lactam antibiotics, can solve the detection needs that are not suitable for low-cost enterprises, the detection sensitivity of colloidal gold test strips is low, and the requirements for detection personnel are high. problems, to achieve the effect of short detection time, easy promotion and use, and wide application range

Active Publication Date: 2012-05-16
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Microbial obstruction detection methods mainly include inhibition zone test, turbidity test, and color-changing detection gold standard kit method. Microbial detection methods are low in cost, but low in sensitivity, time-consuming, and the presence of drug-resistant bacteria may easily lead to false detection; Physical and chemical detection methods are currently the main methods for detecting β-lactam antibiotics, including liquid chromatography, chromatography / mass spectrometry, gas chromatography, etc. The physical and chemical detection methods have high accuracy, but their detection costs are high and the detection equipment Complicated, high requirements for testing personnel, time-consuming and other disadvantages, the currently commonly used immunological analysis methods, mainly enzyme-linked immunoassay and radioimmunoassay, still require high testing costs, which are not suitable for low-cost enterprises detection needs
[0004] Chinese invention patent 200810162588.5 (CN101429241A) discloses "the method and application of penicillin and carrier protein coupling product and β-lactam penicillin antibody preparation", the ELISA kit prepared by the penicillin monoclonal antibody in this method can only To detect three β-lactam antibiotics, penicillin G, ampicillin and carbenicillin, the detection sensitivity of the prepared colloidal gold test strip is low, and there are few types of detection drugs

Method used

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  • Kit and method for detecting beta-lactam antibiotics
  • Kit and method for detecting beta-lactam antibiotics
  • Kit and method for detecting beta-lactam antibiotics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, the composition of the test kit that detects β-lactam antibiotics

[0039] 1. Microporous Reagents

[0040] The micropore reagent has a micropore plug;

[0041] The microporous reagent is freeze-dried with a cephalosporin monoclonal antibody-colloidal gold marker, and the coating amount is 0.20-0.25 μg / ml;

[0042] Two, test paper (called test strip) ( figure 1 ):

[0043] The test paper is composed of a bottom plate, a sample absorption pad, a reaction film, a water absorption pad, and a protective film;

[0044]The sample absorbent pad 1, reaction film 2, water absorbent pad 3 and protective film are pasted on the bottom plate 6 in sequence, the end of the sample absorbent pad is connected to the reaction film, the end of the reaction film is connected to the water absorbent pad, and the sample absorbs The beginning of the pad is aligned with the beginning of the bottom plate, and the end of the absorbent pad is aligned with the end of the bottom pl...

Embodiment 2

[0050] The preparation method of kit described in embodiment 2, embodiment 1

[0051] 1. Preparation of kits

[0052] The preparation method of the kit mainly includes the following steps:

[0053] 1) Prepare freeze-dried microporous reagents with cephalosporin monoclonal antibody-colloidal gold markers;

[0054] 2) preparing a reaction membrane with a detection area coated with a cephalosporin-carrier protein conjugate and a quality control area coated with a goat anti-mouse anti-antibody;

[0055] 3) Assemble the reaction film prepared in 2) with the sample absorbent pad, water absorbent pad, protective film, and bottom plate into a test paper;

[0056] 4) Assemble the freeze-dried microporous reagent and test paper prepared in 1) and 3) with cephalosporin monoclonal antibody-colloidal gold markers into a kit.

[0057] The following step-by-step detailed description:

[0058] (1) Preparation of each component

[0059] 1. Synthesis and identification of cephalosporin-car...

Embodiment 3

[0101] Embodiment 3, the application of test paper

[0102] One, detect β-lactam antibiotics in milk with the kit described in Example 1

[0103] The kit of the invention can detect milk samples. The general detection method is as follows:

[0104] 1. Detection method

[0105] Add the sample solution to be tested dropwise into the microwell reagent, mix well, incubate for 5 minutes, insert the end of the test paper with the MAX line mark down into the microwell reagent, and watch the result within 5 minutes.

[0106] 2. Judgment of test results

[0107] When the concentration of β-lactam antibiotics in the sample is higher than or equal to the minimum detection limit of the kit, the colloidal gold antibody will combine with β-lactam antibiotics, so that it will not bind to the cephalosporin drug conjugate in the detection area due to competitive reaction Combination without a red band is positive. Due to the lack of antibody-antigen competition reaction during the detecti...

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Abstract

The invention discloses a kit and a method for detecting beta-lactam antibiotics. The kit comprises a micropore reagent and a piece of test paper, colloidal gold labeled cephalosporin medicament specific antibodies are freeze-dried in the micropore reagent, and the cephalosporin medicament specific antibodies can be cephalosporin medicament monoclonal antibodies or cephalosporin medicament polyclonal antibodies; the test paper consists of a sample absorption pad, a reaction film, a water absorption pad, a protective film and a bottom plate which are sequentially connected; the reaction film comprises a detection area and a quality control area; the detection area is coated with a cephalosporin medicament-carrier protein conjugate; and the quality control area is coated with anti-antibodies. The method for detecting the beta-lactam antibiotics by using the kit is simple, convenient, fast, intuitive, accurate, wide in application range, low in cost and easy to popularize and use.

Description

technical field [0001] The invention relates to a kit and method for detecting β-lactam antibiotics. Background technique [0002] With the improvement of people's living standards, the drug residues in food of animal origin have been paid more and more attention by the international community. β-lactam antibiotics are the earliest discovered and most widely used antibiotics, and the most reported residues in milk and animal tissues. β-lactam antibiotics refer to a class of antibiotics containing a β-lactam ring in their chemical structure, mainly including penicillins and cephalosporins, and their function is characterized by being able to inhibit the activity of bacterial mucopeptide transpeptidase, thereby preventing bacterial Synthesis of cell walls, exhibits bactericidal activity. In animal production, β-lactam antibiotics are widely used to control mastitis in dairy cows, and treat animal urinary tract, gastrointestinal tract and respiratory tract infections. Howeve...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558
Inventor 何方洋万宇平冯才伟冯静汪善良罗晓琴何丽霞赵正苗冯才茂崔海峰李勇王建霞
Owner BEIJING KWINBON BIOTECH
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